EP0743952A1 - Generation and screening of synthetic drug libraries - Google Patents
Generation and screening of synthetic drug librariesInfo
- Publication number
- EP0743952A1 EP0743952A1 EP95909449A EP95909449A EP0743952A1 EP 0743952 A1 EP0743952 A1 EP 0743952A1 EP 95909449 A EP95909449 A EP 95909449A EP 95909449 A EP95909449 A EP 95909449A EP 0743952 A1 EP0743952 A1 EP 0743952A1
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- EP
- European Patent Office
- Prior art keywords
- containing organic
- organic compounds
- polyhydroxyl
- mixture
- compounds
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/04—Disaccharides
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/02—Acyclic radicals, not substituted by cyclic structures
- C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H5/00—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium
- C07H5/04—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium to nitrogen
- C07H5/06—Aminosugars
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/0068—Means for controlling the apparatus of the process
- B01J2219/00702—Processes involving means for analysing and characterising the products
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00718—Type of compounds synthesised
- B01J2219/0072—Organic compounds
- B01J2219/00731—Saccharides
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- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B40/00—Libraries per se, e.g. arrays, mixtures
- C40B40/04—Libraries containing only organic compounds
- C40B40/12—Libraries containing saccharides or polysaccharides, or derivatives thereof
Definitions
- This invention is concerned with generating libraries of compounds for the discovery of bioactive leads and novel chemical entities.
- the active leads generated and identified by this invention can be used for the development of pharmaceuticals, agrochemicals, and the like.
- Drug discovery processes are very lengthy. A conventional process involves testing and screening of thousands of individual compounds for a desired therapeutical activity. Historically, less than 1 in 10,000 of the synthetic compounds have made it to the drug market, at costs greater than $200 million per drug (Ganellin, C. R., 1992, in Medicinal Chemistry for the 21st Century, Ed. by Wermuth, C. G.; Koga, N.; Konig, H.; and Metcalf, B. W. Blackwell Scientific Publications, London; pp. 3-12).
- Drugs have been sought from natural products for many years. Complex mixtures made from cells or their secondary metabolites have been screened for biological activity. When a desired biological activity has been found in such a complex mixture, the particular chemical which has the activity has been purified, using the biological activity as the means of identifying the component of the mixture which contains the desired activity.
- Carbohydrates are a large family of organic molecules with highly functionalized carbon skeletons. They share the structure C x (H 2 O) y . In fact, this may be the only readily available family of molecules where almost every carbon within the skeleton is functionalized. These functionalizations and related stereochemical differences also result in different chemical reactivities for individual carbon atoms.
- a method for generating and screening a library of compounds comprises: reacting one or more polyhydroxyl-containing organic compounds with subequimolar amounts of displacing groups to form a first mixture comprising displacing group-substituted polyhydroxyl-containing organic compounds; testing said first mixture or a reaction product made therefrom for the presence of a component having a biological activity.
- a method for generating and screening a library of compounds.
- the method comprises: reacting one or more polyhydroxyl-containing organic compounds with subequimolar amounts of protecting groups to form a first mixture of randomly protected polyhydroxyl-containing organic compounds; . oxidizing said first mixture to form a second mixture comprising carbonyl-containing organic compounds;
- Figure 1 represents three reaction schemes for generating a complex mixture of compounds using a mixture of simple saccharides or a single oligosaccharide as the basic starting materials.
- the polyhydroxyl-containing organic compounds used in the present invention have at least two hydroxyl functionalities and contain a carbon-carbon backbone of at least two carbons atoms.
- the carbon-carbon backbone of the compounds may be saturated or unsaturated. cyclic or linear.
- the compounds include: carbohydrates, polyalcohols (e.g., ethylene glycol and glycerol) and polyphenols (e.g., hydroquinones and tetracyclines).
- Carbohydrate, saccharide, sugar and carbohydrate-transformed compounds are defined herein to include ail chemical moieties having a saccharide unit or which are transformed from a saccharide. These terms may also include glycopeptides, glycolipids and other biopolymers (or biomacromolecules) containing saccharides, either in their entirety or as part of the molecular framework.
- polyhydroxylated organic compounds include: monomeric acyclic compounds, such as ethylene glycol, glycerol, and 1, 2, 3,-trihydroxy pentane; polymeric acyclic compounds, such as di- or tri-ethylene diglycol; monomeric cyclic compounds, such as inositol and 1,2,3-trihydroxycyclopentane; polymeric cyclic compounds, such as di-inositol; polymeric and monomeric unsaturated compounds such as tetrahydroxy-1,4- quinone hydrated and polyphenols, such as tetracyclines; and combinations thereof.
- monomeric acyclic compounds such as ethylene glycol, glycerol, and 1, 2, 3,-trihydroxy pentane
- polymeric acyclic compounds such as di- or tri-ethylene diglycol
- monomeric cyclic compounds such as inositol and 1,2,3-trihydroxycyclopentane
- polymeric cyclic compounds such as di-inositol
- a reaction between glycerol and glucose can produce twelve different isomers by glycosidic and anhydride linkages.
- a compound library may also be built via the chemical reactions of phenolics and polyphenolics and other poly oxygenated chemicals. For example, when glycerol is combined via an ether linkage with 2,4-dihydroxytoluene, four different isomers of the toluene- hydroxypropyl ether compounds are made.
- Displacing groups according to the present invention can be any which are known in the art to directly or indirectly displace a hydroxyl group from a carbon atom. These include halogen ions, azide ions, nitriles, sulfites, and alkoxides. Often it is desirable to activate the hydroxyl moieties prior to the step of displacement. Any suitable activation group can be used. Thus activation can be accomplished by reaction with a halide salt, such as bromine or iodine salt, or with methanesulfonyl chloride to yield a sulfonyl ester. Similarly, activation is desirable prior to oligomerizing polyhydroxyl compounds. Useful for such activation reactions are sulfonyl compounds, halide compounds, and thio- compounds.
- the oligomerizing reactions of the present invention include, but are not limited to, (1) the formation of a glycosidic bond, (e.g., the elimination of a water molecule between the anomeric hydroxyl group of one sugar and a hydroxyl group of another sugar); and/or (2) the formation of a sugar anhydride bond, (e.g., the elimination of a water molecule between any hydroxyl group other than those involved in glycosidic bond formation).
- Any reaction condition or sequence that may result in these bond formations can be applied to this synthesis.
- the hydroxyl group will be activated prior to oligomerization. Such conditions are known in the art.
- an anomeric hydroxyl group can be transformed to a halogen functionality (such as CI, Br, or I) when the sugar is treated with an appropriate transition metal salt such as HgX 2 , or the halogen itself, such as X 2 .
- This hydroxyl group can also be transformed into an activated ester-like functionality when treated with toluenesulfonyl or methanesulfonyl chloride.
- reaction products be reduced.
- conditions known in the art for reduction any of which may be used in the practice of the present invention. These include catalytic reduction using Raney- nickel, platinum, palladium, or platinum-carbon. Hydride reduction can also be used. Such reduction may employ LiAlH 4 , NaAlH 4 , and KA1H 4 .
- the multiple hydroxyl functionalities attached to the carbon-carbon backbones can be directly or indirectly transformed into other chemical moieties as desired.
- any of these hydroxyl groups can be displaced randomly by a simple halogen, thereby directly transforming the saccharide(s) into an array of halogenated saccharides.
- these hydroxyl groups may first be activated to sulfonated esters then displaced with either halogen, azide, nitrile, alkoxide, or sulfite, thereby indirectly transforming the saccharide(s) into an array of either halogenated, nitrogenated, sulfurized, and/or oligomerized saccharides
- the saccharide may also be transformed into other chemical moieties indirectly via oxidation of the hydroxyl functionalities.
- the hydroxyl functionalities of saccharides can be oxidized to carbonyl functionalities randomly to give a family of partially oxidized sugars. These partially oxidized sugars can subsequently be transformed into other chemical functionalities based on the reactions of the carbonyls, thereby indirectly transforming the saccharides into an array of new chemical entities which may not resemble the starting material.
- Prior to oxidizing hydroxyl functionalities it is desirable to react the polyhydroxyl- containing organic compounds with subequimolar amounts of protecting groups. Protecting groups are those which are not sensitive to the oxidizing conditions used to transform the hydroxyl groups to carbonyl moieties.
- a combination of such functional groups can be introduced directly or indirectly so that any number of the original hydroxyl groups of a saccharide are displaced by other chemical moieties.
- the oligomerization and functionalization of polyhydroxyl compounds such as saccharides can be carried out sequentially or concurrently.
- the number of organic compounds present in each library, as a consequence of prior chemical events, will be multiplied by the next random chemical transformation to expand the collection of organic compounds.
- the present invention it is desirable to minimize the differences among saccharidic hydroxyl groups during the reaction process, and thus force the formation of multiple reaction products. This is contrary to the conventional strategy in synthetic chemistry, where the purpose of the reaction is to form a particular product, and reaction specificity is the aim. However, the goal of this invention is to provide means to generate many reaction products, rather than to identify specific reaction conditions where a limited number of carbohydrate derivatives can be generated. According to the present invention differences among saccharidic hydroxyl groups are minimized by using small and highly reactive reagents to minimize the steric hindrance during the reaction process.
- Such reagents include methanesulfonyl chloride, toluenesulfonyl chloride, and halogen salts such as PBr 5 , PBr 3 , HBr, HI, and Lil.
- Other means for reducing reaction specificity include: (1) selectively blocking the reaction sites that are either more reactive than the others (such as the (C-l anomeric hydroxyl groups) or sterically more accessible than the others (such as the C-6 hydroxyl group of a hexopyranoside); and/or (2) using sub-molar equivalent (subequimolar) amounts
- protecting groups as are known in the art.
- protecting groups include CH 3 O . ?nd C J H J O " , etc., which can be derived from the a nols.
- groups include trityl (triphenylmethyl), TPS (triphenylsilyl), TBMPS (t- butylmethoxyphenylsilyl), etc.
- Useful protecting groups are those which are not displaced by the displacing groups which are employed in the subsequent step.
- the present invention employs a multiplicity of linkages between monomers, large numbers of sequences of linked monomers, and a multiplicity of different chemical transformation products.
- This invention makes use of highly functionalized polyhydroxylated compounds ("polyhydroxyl-containing organic compounds") to generate chemical libraries.
- polyhydroxyl-containing organic compounds highly functionalized polyhydroxylated compounds
- Libraries which are generated can be screened for any biological activity known in the art. These include, but are not limited to, antimicrobial activity, antitumor activity, enzyme inhibiting activity, receptor binding, growth promotion activity, and in vitro and in vivo tests for biological responses. Many assays are known for these activities in the art and any can be used as is convenient.
- fractionation of the mixture or library is desirable. Any fractionating means known in the art can be employed. Typically these will involve solvent partitioning and/or conventional chromatography, which may include liquid column chromatography, thin layer chromatography, high performance liquid chromatography, affinity chromatography, or ion exchange chromatography. Individual fractions can then be tested to locate the fraction with the desired biological activity. Fractionation can be repeated or combined with other fractionation steps and the fractions again tested for activity. In such a manner, the active ingredient can be isolated and purified from the highly complex mixture of reaction products.
- Figure 1 summarizes the synthetic schemes used to generate the compound libraries.
- Table 1 summarizes the antimicrobial assay results of these libraries against methicillin resistant Stc ⁇ hylococcus aureus (MRS A), Bacillus subtilis (B. sub), Pseudomonas aeruginosa (Ps. aerug.), E. coli, and Candida albicans (C. alb.). Synthetic methods are given in detail in the examples. The in vitro bacterial susceptibility tests were conducted according to the method specified in
- Examples I - III depicted in Figure 1, and described in more detail below a sequence of chemical libraries was built, one upon the other, starting with three simple monosaccharides, namely glucose, galactose, and inositol (in 1:1:1 ratio).
- methanesulfonyl chloride was introduced to a mixture of the three monosaccharides under a basic condition (pyridine, a basic solvent).
- the resultant sulfonyl esters generated in situ activated the hydroxyl functionalities of the sugars.
- These activated ester moieties could be either displaced by the hydroxyl group of another sugar, which would produce either oligomeric glycosides or sugar anhydrides, or displaced by another nucleophilic group, such as azide.
- library #1 contained deoxy-azido-sugars in either oligomeric or monomeric forms.
- TSK-1000 HXL indicated that there were nearly 14% dimers present in library #1.
- the oligomerization reaction introduced here significantly increased the possible number of different oligomers present in library #2 by formation of different glycosidic linkages.
- a portion of library #1 was treated with Hg(CN) 2 , HgBr 2 and Hgl 2 in acetonitrile in the presence of base to increase the percentage of the oligomeric sugars, which led to the expanded library #2.
- This reaction step expanded the collection of different oligomers via the anomeric carbon (C-l) of the sugars. As indicated by the same gel permeation analysis, 27% dimers and trimers were found (indicating a 13% increase in the oligomers).
- the deoxy-azido-sugar library #2 was converted to the deoxy-amino-sugar library #3 via catalytic reduction using hydrogen and palladium oxide on carbon. This reaction did not appreciably increase the number of compounds present in the library; rather, it converted the compounds from azido- functionality to the corresponding amino- functionality.
- the resultant library #3 was partitioned between water and ethyl acetate to give library #3a, the aqueous fraction of library #3, and library #3b, the organic fraction of library #3.
- Table 1 summarizes the antimicrobial activity profile of libraries #1, #2, #3a and #3b. Note that all of the libraries showed antimicrobial activity.
- the deoxy-azido sugar libraries #1 and #2 showed activity only against gram positive bacteria, B. subtilis and Staphylococcus aureus.
- the deoxy-amino sugar library #3a showed activities against the same species of gram-positive bacteria as well as the gram-negative bacteria, E. coli and Pseudomonas aeruginosa.
- the organic fraction, library #3b only showed activity against B. subtilis .
- Examples IV - VII depicted in Figure 1 used a single trisaccharide, raffinose 1 , to prepare a sequence of libraries containing aminoglycosides.
- the azide-library #4 was then converted to an amino-sugar library by catalytic reduction, and the benzylformate protecting groups were removed to produce the amino-sugar library #5.
- the resultant library #5 was partitioned between organic solvent and water to give libraries #5a, the aqueous fraction, and #5b, the organic fraction.
- Examples VI and VII which produced libraries #6, #7a, #7b, and #7c, started with the same starting material as libraries #4 and #5, and used the same reaction sequence to introduce nitrogen and generate amino-sugars.
- Examples VI and VII differ from IV and V in that different hydroxyl protecting groups were introduced.
- 2 molar equivalents of CH 3 I were introduced so that an average of two raffinose hydroxyl groups were converted to a methoxyl functionality.
- 4 molar equivalents of benzyl group were introduced, so an average of 4 hydroxyl groups per molecule would not be converted or displaced by other chemical functionalities, and could be converted back to free hydroxyl upon catalytic reduction.
- library #6 was then converted to the deoxy-azido-sugar library #7a, followed by catalytic reduction and partition to give libraries #7b, the aqueous fraction, and #7c, the organic fraction.
- libraries differ in the means used to introduce the different hydroxyl protecting groups and the methylation of the aminoglycoside libraries.
- libraries #4 - #5 a benzylformate protecting group was used, whereas the other libraries used a mixture of methyl and benzyl protecting groups.
- library #7b was randomly methylated whereas the library #5a was not.
- Library #7b exhibited anti-yeast (C. albicans) activity, whereas the library #5 exhibited antibacterial activity, as indicated in Table 1.
- the aqueous layer was freeze-dried to give a crude solid.
- the proton NMR analysis indicated that this fraction is the pyridinium methanesulfonate salt.
- the successful displacement is first indicated by the changing of the carbohydrate solubility from polar protic solvent (H 2 O) to polar aprotic solvent (CH 3 CN).
- TSK 1000 HXL/THF LC and HPLC gel permeation
- the column was then developed with HsOH/CH ⁇ N (50:50) in which the crude product was dissolved. A total of 11 fractions (ca. 75 ml each) were collected. The sugar-containing fractions (frac. 2-6) indicated by NMR analysis were combined and the solvents were removed by vacuum to give library #2, a light yellow colored solid (376.8 mg, ca. 60% of the crude mixture).
- the HPLC gel permeation analysis of the crude mixture indicated that the percentage of the oligomerization product with molecular weight about 400 increased from 14% to 27%.
- the biological activity profiles of the library #2 is shown in Table 1.
- reaction mixture was then stirred under the above conditions for another 30 min. followed with the addition of an additional portion of methanesulfonyl chloride (0.4 mL).
- the entire reaction mixmre was gradually warmed up to ambient temperature over 45 mins. and then allowed to stir under Ar for another 16 hrs.
- the pyridinium salt precipitate was then removed by filtration (widi a double-end filter), and die filtrate was directly transferred into a flask containing NaN 3 (600 mg).
- the reaction mixmre was men stirred at room temperature under Ar for 72 hr.
- the solvent was removed by vacuum (freeze dried over night); the resultant solid was partitioned between 100 mL of 10% MeOH in ethyl acetate (v:v) and water (eq.
- a portion of Library #4 (ca. 700 mg) was suspended in a mixture of organic solvents (ethylacetate and memanol, 1:1, v;v; 50 mL), to which a catalytic amount of palladium oxide on carbon (50 mg) was added. The solution was then purged with hydrogen for 4 hrs with vigorous stirring at ambient temperature. The reaction vessel was sealed under a positive pressure of hydrogen (2-5 psi) and stirred at ambient temperature overnight. The reaction slurry was then filtered, and the catalyst washed with ethyl acetate, memanol and water. All filtrates and washes were combined and concentrated to dryness to give a white amorphous solid.
- organic solvents ethylacetate and memanol, 1:1, v;v; 50 mL
- the resultant oily solid (light tan colored) was suspended in a methanol/water (50/50, v:v) solution, and freeze-dried again to force a complete removal of the solvent.
- the dried solid was suspended in denatured etiianol (100 mL) and the insoluble residue was filtered to give a clear filtrate.
- the filtrate was then concentrated to dryness (9.0 grams). Analysis by reverse phase thin layer chromatography and NMR spectroscopy indicated the formation of multiple sugar- containing products.
- the crude mixmre was then loaded onto a Toyopearl 40F gel permeation column and eluted with 5% (v.v) methanol in water.
- the solution was then stirred vigorously under Ar at ambient temperature for 16 hrs.
- the reaction temperamre was brought to 50° C with an oil-bath under Ar for 2 additional hrs, and cooled to die ambient temperature before filtration.
- the filtrate was then freeze-dried to remove me pyridine.
- the resulting oily solid was suspended in a mixmre of solvents (H 2 O, 3 mL; MeOH, 20 mL; and EtOAc, 10 mL).
- the milky suspension was loaded onto Toyopearl 40F column which was men eluted wid EtOAc (100%).
- the fractions containing the OBz (broad multiplets at 7.1 to 7.5 ppm.
- the solid was tiien partitioned between 250 mL of water and 250 mL of EtOAc. The layers was then separated. The organic layer was washed once again with water, while the aqueous layer was washed once widi EtOAc. The washes were then combined widi the corresponding layers of extracts and solutions were then concentrated to dryness. The aqueous fraction gave a clear oily solid Gibrary #7b, 194 mg) whereas the organic layer gave a light yellow colored solid (library #7c, 176 mg).
- the biological activity test results of the libraries #7b and 7c are given in Table 1.
Abstract
Methods are provided for generating highly diverse mixtures of compounds which may be screened for biological activities. Once the activity is found, the component of the mixture which is responsible for the activity can be isolated by fractionation and assay for the biological activity. Polyhydroxylated organic monomers and oligomers are used as starting materials for generating the libraries.
Description
GENERATION AND SCREENING OF SYNTHETIC DRUG LIBRARIES
TECHNICAL FIELD OF THE INVENTION
This invention is concerned with generating libraries of compounds for the discovery of bioactive leads and novel chemical entities. The active leads generated and identified by this invention can be used for the development of pharmaceuticals, agrochemicals, and the like. BACKGROUND OF THE INVENTION
Drug discovery processes are very lengthy. A conventional process involves testing and screening of thousands of individual compounds for a desired therapeutical activity. Historically, less than 1 in 10,000 of the synthetic compounds have made it to the drug market, at costs greater than $200 million per drug (Ganellin, C. R., 1992, in Medicinal Chemistry for the 21st Century, Ed. by Wermuth, C. G.; Koga, N.; Konig, H.; and Metcalf, B. W. Blackwell Scientific Publications, London; pp. 3-12).
Drugs have been sought from natural products for many years. Complex mixtures made from cells or their secondary metabolites have been screened for biological activity. When a desired biological activity has been found in such a
complex mixture, the particular chemical which has the activity has been purified, using the biological activity as the means of identifying the component of the mixture which contains the desired activity.
An alternative method for screening compounds for desirable biological
activities has been to screen individual compounds which have been synthesized and saved in libraries of drug or chemical companies or research institutes. The compounds in these libraries were often chosen for synthesis and screening because they had a particular functionality thought to be relevant to a particular biological activity.
More recently, some companies have begun to create their own peptide and oligonucleotide libraries to screen for a particular biological function. "Peptide Library Synthesis and Screening Strategies for Drug Development", Genetic Engineering News, May 1, 1993, page 6; "Ixsys Licenses In Vitro Monoclonal Process", Genetic Technology News, vol. 12, page 14, October 1992; and Ladner, U.S. Patent 5,223,409; "In Vitro Evolution Creates Novel Drugs," Genetic Engineering News, page 1, April 15, 1993. The combinatorial peptide and nucleotide libraries described to date involve the sequence randomization of individual monomers using a single naturally existing biological linkage (such as 3 '-5' phosphate linkage of nucleotides or amide linkage of peptides).
J.H. Musser, "Trends in New Lead Identification" in Medicinal Chemistry for the 21st Century, edited by Wermuth, Koga, Konig and Metcalf. teaches that carbohydrates have the potential for greater complexity than polypeptides or
oligonucleotides. He opines that "of all the structural types, carbohydrates have the greatest theoretical potential for specificity and new lead generation. "
Carbohydrates are a large family of organic molecules with highly functionalized carbon skeletons. They share the structure Cx(H2O)y. In fact, this may be the only readily available family of molecules where almost every carbon within the skeleton is functionalized. These functionalizations and related stereochemical differences also result in different chemical reactivities for individual carbon atoms.
The great potential of carbohydrates for use in generating organic compound libraries rests within the inherent character of this class of compounds. Almost every carbon in a given carbohydrate has a hydroxyl functionality (or other oxygen-containing functional group) attached to it. Different spatial Λgements (stereochemical arrangements) of these hydroxyl groups result in different carbohydrates; for example, the difference between glucose and galactose is attributable i the stereochemistry of the hydroxyl group attached to the < carbon. Different linkages between carbohydrates derived from these hydroxyl functionalities (i.e., by removal of a water molecule) also result in different carbohydrates. As a consequence, the potential number of different saccharides generated by combining several saccharides together can be very large.
When two hexoses are linked together via a glycosidic bond (a chemical linkage involving the anomeric carbon of at least one of the saccharides), eleven different disaccharides can theoretically be obtained. A mixture of thref monosaccharides can, in theory, generate as many as 1056 possible trisaccharide combinations, and a mixture of five different monosaccharides can be assembled
into 31 million pentasaccharides. In addition, if the reaction conditions produce sugar anhydride linkages, the combinatory possibilities increase many fold. For instance, two identical monosaccharides can generate a total number of 26 different dimers of sugar anhydrides and glycosides.
Recently, new screening processes have been developed which have the potential for high throughput, i.e., many compounds per unit time can be tested individually for a particular biological activity. See "Progenies Improves the Screening Process", Genetic Technology News, vol. 9, page 5, May 1989; "Nova gets $2.5 Million for AIDS", Biotechnology Newswatch, vol. 8, page 2, December 19, 1988; "RT speeds screening for new drugs" Chemical Week, April 15, 1987, page 19. Such screening techniques involve testing for inhibitors of specific proteins, or binding to particular cellular receptors. These screening processes are typically performed on pre-existing libraries of chemicals. See "Newsfront: Companies" Chemical Engineering, page 35, September 16, 1985; and "RT speeds screening for new drugs" Chemical Week, April 15, 1987, page 19.
The potential diversity of polyhydroxyl-containing organic compounds and the newly developed screening procedures, have together generated an opportunity for devising new methods to generate rapidly novel, large libraries of diverse chemicals for screening for bioactive compounds.
SUMMARY OF THE INVENTION
It is an object of the invention to provide methods for generating libraries of compounds.
It is another object of the invention to provide a method to rapidly produce large libraries of diverse compounds.
It is still another object of the invention to provide a synthetic strategy for the chemical preparation of a library of compounds based on polyhydroxyl- contøining organic compounds.
It is yet another object of the invention to provide a library of compounds by means of chemical synthesis.
It is another object of the invention to provide a library of compounds for use m pharmaceutical and agrochemical screening and discovery.
It is still another object of the invention to generate compounds for therapeutic use.
These and other objects of the invention are provided by one or more of the embodiments described below. In one embodiment a method for generating and screening a library of compounds is provided which comprises: reacting one or more polyhydroxyl-containing organic compounds with subequimolar amounts of displacing groups to form a first mixture comprising displacing group-substituted polyhydroxyl-containing organic compounds; testing said first mixture or a reaction product made therefrom for the presence of a component having a biological activity.
According to a second embodiment of the invention a method is provided for generating and screening a library of compounds. The method comprises: reacting one or more polyhydroxyl-containing organic compounds with subequimolar amounts of protecting groups to form a first mixture of randomly protected polyhydroxyl-containing organic compounds; .
oxidizing said first mixture to form a second mixture comprising carbonyl-containing organic compounds;
testing said first or said second mixtures or a reaction product made therefrom for the presence of a component having a biological activity.
These and other embodiments of the invention, which are described in more detail below, provide the art with means of generating complex mixtures of compounds which may or may not exist in nature and of determining useful compounds among said complex mixtures. BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 represents three reaction schemes for generating a complex mixture of compounds using a mixture of simple saccharides or a single oligosaccharide as the basic starting materials. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
It is a discovery of the present invention that complex mixtures of polyhydroxyl compounds can be generated, transformed, and screened in large numbers by means of randomizing chemosynthetic strategies. Using such strategies, useful compounds have been synthesized and identified in pools of compounds.
The polyhydroxyl-containing organic compounds used in the present invention have at least two hydroxyl functionalities and contain a carbon-carbon backbone of at least two carbons atoms. The carbon-carbon backbone of the compounds may be saturated or unsaturated. cyclic or linear. The compounds include: carbohydrates, polyalcohols (e.g., ethylene glycol and glycerol) and polyphenols (e.g., hydroquinones and tetracyclines). Carbohydrate, saccharide,
sugar and carbohydrate-transformed compounds are defined herein to include ail chemical moieties having a saccharide unit or which are transformed from a saccharide. These terms may also include glycopeptides, glycolipids and other biopolymers (or biomacromolecules) containing saccharides, either in their entirety or as part of the molecular framework.
Carbohydrates, though promising as Musser pointed out, and discussed extensively herein for the purpose of demonstration, merely represent a portion of a much larger family of polyhydroxylated organic compounds useful for purposes of this invention. In addition to carbohydrates, polyhydroxylated organic compounds include: monomeric acyclic compounds, such as ethylene glycol, glycerol, and 1, 2, 3,-trihydroxy pentane; polymeric acyclic compounds, such as di- or tri-ethylene diglycol; monomeric cyclic compounds, such as inositol and 1,2,3-trihydroxycyclopentane; polymeric cyclic compounds, such as di-inositol; polymeric and monomeric unsaturated compounds such as tetrahydroxy-1,4- quinone hydrated and polyphenols, such as tetracyclines; and combinations thereof. For example a reaction between glycerol and glucose can produce twelve different isomers by glycosidic and anhydride linkages. A compound library may also be built via the chemical reactions of phenolics and polyphenolics and other poly oxygenated chemicals. For example, when glycerol is combined via an ether linkage with 2,4-dihydroxytoluene, four different isomers of the toluene- hydroxypropyl ether compounds are made.
Key to the achievement of randomization of reaction products is the use of subequimolar amounts of reactants, either displacing groups or protecting groups. Use of such amounts maximizes the number of different products which can be
achieved. Equimolar amounts, according to the present invention, are assessed against the number of hydroxyl moieties which are available for reaction.
Displacing groups according to the present invention can be any which are known in the art to directly or indirectly displace a hydroxyl group from a carbon atom. These include halogen ions, azide ions, nitriles, sulfites, and alkoxides. Often it is desirable to activate the hydroxyl moieties prior to the step of displacement. Any suitable activation group can be used. Thus activation can be accomplished by reaction with a halide salt, such as bromine or iodine salt, or with methanesulfonyl chloride to yield a sulfonyl ester. Similarly, activation is desirable prior to oligomerizing polyhydroxyl compounds. Useful for such activation reactions are sulfonyl compounds, halide compounds, and thio- compounds.
The oligomerizing reactions of the present invention include, but are not limited to, (1) the formation of a glycosidic bond, (e.g., the elimination of a water molecule between the anomeric hydroxyl group of one sugar and a hydroxyl group of another sugar); and/or (2) the formation of a sugar anhydride bond, (e.g., the elimination of a water molecule between any hydroxyl group other than those involved in glycosidic bond formation). Any reaction condition or sequence that may result in these bond formations can be applied to this synthesis. Typically the hydroxyl group will be activated prior to oligomerization. Such conditions are known in the art. For example, an anomeric hydroxyl group can be transformed to a halogen functionality (such as CI, Br, or I) when the sugar is treated with an appropriate transition metal salt such as HgX2, or the halogen itself, such as X2. This hydroxyl group can also be transformed into an activated ester-like
functionality when treated with toluenesulfonyl or methanesulfonyl chloride. These resultant products from the above reactions can readily react with the hydroxyl functionalities of another sugar to give a glycosidic linkage. These transformations can also occur at the hydroxyl groups other than the one directly attached to th anomeric carbon, therefore leading to the formation of sugar anhydrides. Oligomerizing reactions, according to the present invention, are a particular type of the class of displacement reactions in which the displacing group is an alkoxide.
Often it is desirable that the reaction products be reduced. There are many sets of conditions known in the art for reduction, any of which may be used in the practice of the present invention. These include catalytic reduction using Raney- nickel, platinum, palladium, or platinum-carbon. Hydride reduction can also be used. Such reduction may employ LiAlH4, NaAlH4, and KA1H4.
The multiple hydroxyl functionalities attached to the carbon-carbon backbones can be directly or indirectly transformed into other chemical moieties as desired. For instance, any of these hydroxyl groups can be displaced randomly by a simple halogen, thereby directly transforming the saccharide(s) into an array of halogenated saccharides. In addition, these hydroxyl groups may first be activated to sulfonated esters then displaced with either halogen, azide, nitrile, alkoxide, or sulfite, thereby indirectly transforming the saccharide(s) into an array of either halogenated, nitrogenated, sulfurized, and/or oligomerized saccharides
The saccharide may also be transformed into other chemical moieties indirectly via oxidation of the hydroxyl functionalities. For example, the hydroxyl
functionalities of saccharides can be oxidized to carbonyl functionalities randomly to give a family of partially oxidized sugars. These partially oxidized sugars can subsequently be transformed into other chemical functionalities based on the reactions of the carbonyls, thereby indirectly transforming the saccharides into an array of new chemical entities which may not resemble the starting material. Prior to oxidizing hydroxyl functionalities it is desirable to react the polyhydroxyl- containing organic compounds with subequimolar amounts of protecting groups. Protecting groups are those which are not sensitive to the oxidizing conditions used to transform the hydroxyl groups to carbonyl moieties.
A combination of such functional groups can be introduced directly or indirectly so that any number of the original hydroxyl groups of a saccharide are displaced by other chemical moieties. The oligomerization and functionalization of polyhydroxyl compounds such as saccharides can be carried out sequentially or concurrently. The number of organic compounds present in each library, as a consequence of prior chemical events, will be multiplied by the next random chemical transformation to expand the collection of organic compounds.
According to the present invention it is desirable to minimize the differences among saccharidic hydroxyl groups during the reaction process, and thus force the formation of multiple reaction products. This is contrary to the conventional strategy in synthetic chemistry, where the purpose of the reaction is to form a particular product, and reaction specificity is the aim. However, the goal of this invention is to provide means to generate many reaction products, rather than to identify specific reaction conditions where a limited number of carbohydrate derivatives can be generated. According to the present invention
differences among saccharidic hydroxyl groups are minimized by using small and highly reactive reagents to minimize the steric hindrance during the reaction process. Such reagents include methanesulfonyl chloride, toluenesulfonyl chloride, and halogen salts such as PBr5, PBr3, HBr, HI, and Lil. Other means for reducing reaction specificity include: (1) selectively blocking the reaction sites that are either more reactive than the others (such as the (C-l anomeric hydroxyl groups) or sterically more accessible than the others (such as the C-6 hydroxyl group of a hexopyranoside); and/or (2) using sub-molar equivalent (subequimolar) amounts
of chemical reagents in the transformation steps. Selective blocking can be accomplished by using protecting groups as are known in the art. For the hydroxyl groups on the anomeric carbon, such groups include CH3O . ?nd CJHJO", etc., which can be derived from the a nols. For the stereochemically most accessible C-6 site of a hexose pyranoside, such as glucose pyranoside, such groups include trityl (triphenylmethyl), TPS (triphenylsilyl), TBMPS (t- butylmethoxyphenylsilyl), etc. Useful protecting groups are those which are not displaced by the displacing groups which are employed in the subsequent step.
In contrast to the combinatorial peptide and nucleotide libraries described to date which involve the sequence randomization of individual monomers using a single, naturally existing, biological linkage, the present invention employs a multiplicity of linkages between monomers, large numbers of sequences of linked monomers, and a multiplicity of different chemical transformation products. This invention makes use of highly functionalized polyhydroxylated compounds ("polyhydroxyl-containing organic compounds") to generate chemical libraries.
The members of this collection will include the naturally existing biological linkages among other linkages.
Libraries which are generated can be screened for any biological activity known in the art. These include, but are not limited to, antimicrobial activity, antitumor activity, enzyme inhibiting activity, receptor binding, growth promotion activity, and in vitro and in vivo tests for biological responses. Many assays are known for these activities in the art and any can be used as is convenient.
Once a mixture or library has been found which contains a desired biological activity, fractionation of the mixture or library is desirable. Any fractionating means known in the art can be employed. Typically these will involve solvent partitioning and/or conventional chromatography, which may include liquid column chromatography, thin layer chromatography, high performance liquid chromatography, affinity chromatography, or ion exchange chromatography. Individual fractions can then be tested to locate the fraction with the desired biological activity. Fractionation can be repeated or combined with other fractionation steps and the fractions again tested for activity. In such a manner, the active ingredient can be isolated and purified from the highly complex mixture of reaction products.
In the following examples we demonstrate that: (1) a mixture of three common monosaccharides, through a sequence of chemical transformations, can be used to generate a mixture of aminoglycosides and their analogs, some of which are active against pathogens (Fig. la and Table 1, libraries #1 to #3a and #3b); and (2) the products of a single trisaccharide are active against infectious
pathogens after a sequence of chemical transformations (Fig. lb, c and Table 1, libraries #4-7c).
Figure 1 summarizes the synthetic schemes used to generate the compound libraries. Table 1 summarizes the antimicrobial assay results of these libraries against methicillin resistant Stcφhylococcus aureus (MRS A), Bacillus subtilis (B. sub), Pseudomonas aeruginosa (Ps. aerug.), E. coli, and Candida albicans (C. alb.). Synthetic methods are given in detail in the examples. The in vitro bacterial susceptibility tests were conducted according to the method specified in
the 4th edition oi Manual of Clinical Microbiology (Barry, A.L.; and Thornsberry,
C. 1985. Ed. by Edwin H. Lennette (editor-in-chief). American Society for Microbiology, Washington, D.C.; pp. 978).
In Examples I - III depicted in Figure 1, and described in more detail below, a sequence of chemical libraries was built, one upon the other, starting with three simple monosaccharides, namely glucose, galactose, and inositol (in 1:1:1 ratio). First, methanesulfonyl chloride was introduced to a mixture of the three monosaccharides under a basic condition (pyridine, a basic solvent). The resultant sulfonyl esters generated in situ activated the hydroxyl functionalities of the sugars. These activated ester moieties could be either displaced by the hydroxyl group of another sugar, which would produce either oligomeric glycosides or sugar anhydrides, or displaced by another nucleophilic group, such as azide. Due to the multiple hydroxyl functionalities of sugars, both chemical processes (i.e., the oligomerizations by other sugar displacements of the activated esters and the displacements by azide) could happen simultaneously. As a
consequence of the randomized reactions, library #1 contained deoxy-azido-sugars in either oligomeric or monomeric forms. HPLC gel permeation analysis using
TSK-1000 HXL indicated that there were nearly 14% dimers present in library #1.
The oligomerization reaction introduced here significantly increased the possible number of different oligomers present in library #2 by formation of different glycosidic linkages. A portion of library #1 was treated with Hg(CN)2, HgBr2 and Hgl2 in acetonitrile in the presence of base to increase the percentage of the oligomeric sugars, which led to the expanded library #2. This reaction step expanded the collection of different oligomers via the anomeric carbon (C-l) of the sugars. As indicated by the same gel permeation analysis, 27% dimers and trimers were found (indicating a 13% increase in the oligomers).
Following the library expansion reactions, the deoxy-azido-sugar library #2 was converted to the deoxy-amino-sugar library #3 via catalytic reduction using hydrogen and palladium oxide on carbon. This reaction did not appreciably increase the number of compounds present in the library; rather, it converted the compounds from azido- functionality to the corresponding amino- functionality. The resultant library #3 was partitioned between water and ethyl acetate to give library #3a, the aqueous fraction of library #3, and library #3b, the organic fraction of library #3.
Table 1 summarizes the antimicrobial activity profile of libraries #1, #2, #3a and #3b. Note that all of the libraries showed antimicrobial activity. The deoxy-azido sugar libraries #1 and #2 showed activity only against gram positive bacteria, B. subtilis and Staphylococcus aureus. whereas the deoxy-amino sugar library #3a showed activities against the same species of gram-positive bacteria as
well as the gram-negative bacteria, E. coli and Pseudomonas aeruginosa. The organic fraction, library #3b, only showed activity against B. subtilis .
As discussed previously, one can establish a library of chemical compounds by randomly displacing the hydroxyl functionality with another chemical moiety, such as a nitrogen containing group. Examples IV - VII depicted in Figure 1 used a single trisaccharide, raffinose1, to prepare a sequence of libraries containing aminoglycosides.
In Example IV, raffinose, which contains eleven free-hydroxyl groups, was
first reacted with four molar equivalents of benzylchloroformate in pyridine. Introduction of die benzylformate protecting group ensured that an average of 4 molar equivalents of hydroxyl functionality were present in the subsequent amino-sugar libraries. Immediately following the random protecting steps, 7 molar equivalents of methanesulfonyl chloride were introduced to generate the activated esters in situ, which were then converted to a deoxy-azido-sugar library #4 by azide displacement.
The azide-library #4 was then converted to an amino-sugar library by catalytic reduction, and the benzylformate protecting groups were removed to produce the amino-sugar library #5. The resultant library #5 was partitioned between organic solvent and water to give libraries #5a, the aqueous fraction, and #5b, the organic fraction.
Libraries #4, #5a and #5b were tested against a battery of microbes. The results are shown in Table 1. Library #4 did not exhibit any antimicrobial
l3P- TI »-fructofuranosy 1-O-α-D-galactopyranosyH 1 -6)-α-D-glucopyranoside
activity, whereas library #5a showed activity against gram positive bacteria and #5b showed a limited activity against E. coli.
Examples VI and VII, which produced libraries #6, #7a, #7b, and #7c, started with the same starting material as libraries #4 and #5, and used the same reaction sequence to introduce nitrogen and generate amino-sugars. Examples VI and VII differ from IV and V in that different hydroxyl protecting groups were introduced. First, 2 molar equivalents of CH3I were introduced so that an average of two raffinose hydroxyl groups were converted to a methoxyl functionality. Following this reaction, 4 molar equivalents of benzyl group were introduced, so an average of 4 hydroxyl groups per molecule would not be converted or displaced by other chemical functionalities, and could be converted back to free hydroxyl upon catalytic reduction. These two consecutive alkylation steps led to the formation of library #6. As in all of the previous reaction schemes, library #6 was then converted to the deoxy-azido-sugar library #7a, followed by catalytic reduction and partition to give libraries #7b, the aqueous fraction, and #7c, the organic fraction.
The biological activities of libraries #6, #7a, #7b and fflc are indicated in Table 1. Libraries #6, #7a, and #7c did not exhibit any antimicrobial activity against the test organisms. Library #7b showed activity against E. coli and Candida albicans.
As demonstrated by these examples, different spectra of biological activities were obtained when the preparation of compound libraries started with different saccharides. As shown in Table 1, libraries #1 - #3 exhibited a broader spectrum of biological activity against the battery of chosen microbes than did libraries #4
- #7. The primary difference was that the preparation of compound libraries #1
- #3 started with three different monosaccharides. The aminoglycoside library #3a exhibited a broad spectrum of antimicrobial activity against both gram positive and gram negative bacteria. On the other hand, the preparation of libraries #4 - #7 started with a single trisaccharide to give a sequence of aminoglycoside libraries. These libraries exhibited different antimicrobial profiles, as indicated in Table 1.
Starting with the same material, different chemical L. ^raries can be prepared with different biological activity profiles. As demonstrated in Table 1, the
compound libraries #4 - #5 and #6 - #7 were prepared with raffinose. These
libraries differ in the means used to introduce the different hydroxyl protecting groups and the methylation of the aminoglycoside libraries. In the sequence which gave libraries #4 - #5, a benzylformate protecting group was used, whereas the other libraries used a mixture of methyl and benzyl protecting groups. After the catalytic reduction, library #7b was randomly methylated whereas the library #5a was not. Library #7b exhibited anti-yeast (C. albicans) activity, whereas the library #5 exhibited antibacterial activity, as indicated in Table 1.
Examples
Example I.
Preparation of a saccharide based organic compound Library #1.
A mixture of glucose (2 g, 11.11 mM), galactose (2 g, 11.11 mM) and inositol (2 g, 11.11 mM) was suspended in freshly distilled (over CaH,) pyridine (ca. 100 mL) and stirred at room temperature (25° C) for 16 hrs under Ar. Methanesulfonyl chloride (14 mL, 5.34 Meq) was added to the reaction mixture
dropwise under Ar while the reaction mixture was chilled with an ice bath. The reaction temperature was then gradually brought back to ambient temperature as
the ice melted (approx. 45 min). The resultant solution was then stirred vigorously under Ar at ambient temperature for another 16 Hrs. The resultant pyridinium chloride precipitate was then removed via filtration with a double-end schlenck filter under reduced pressure and a positive flow of Ar. The filtrate was directly transferred over solid sodium azide (6.6 g, 3 Meq). The suspension was then stirred at ambient temperature for another 48 hrs under Ar. The reaction mixture was then frozen with liquid N2, and left on the vacuum ( ca. 0.5 - 1 torr) until the pyridine was completely evaporated (ca. 16-24 hrs) to give a dark-brown colored amorphous solid. A mixture of distilled and deionized H2O and acetonitrile (70 mL : 100 mL) was added to the solid and die mixture was left to stand at room temperature for about 10 min. HC1 (0.1%, aq, 200 uL) was then added to the solution, which immediately separated into two layers. The aqueous layer (bottom) was then separated from the organic layer (top) using a separatory funnel. The aqueous layer was then washed twice with acetonitrile (100 mL x 2). The organic extract and washes were then combined and concentrated under vacuum with rotary-evaporator and vacuum (0.5 to 1 torr) to give a brown-colored amorphous solid as the crude mixture of library #1 (19.65 grams). The aqueous layer was freeze-dried to give a crude solid. The proton NMR analysis indicated that this fraction is the pyridinium methanesulfonate salt. The successful displacement is first indicated by the changing of the carbohydrate solubility from polar protic solvent (H2O) to polar aprotic solvent (CH3CN). The LC and HPLC gel permeation (TSK 1000 HXL/THF) analysis indicated that approximately 14%
of the crude mixmre were the products of dimerization (M.W. > 400). The presence of the multiple N3 on the carbohydrate skeletons and their different locations is indicated by the same analysis. Their different substitution location on the carbohydrate skeletons can also be observed. The presence of the proton signal from aldehydes rather than from the anomeric carbon is due to the azide displacement at the C-4 and C-5 position of the carbohydrate skeleton. This displacement trapped the sugars in their aldehyde form rather than their common pyranoside and furanoside form. The mixture was then tested for in vitro antimicrobial activities using the impregnated disc-assay. The biological activity test results of library #1 are given in Table 1.
Example II.
Expansion of the saccharide Library #1 to Library #2.
A portion of the crude library #1 (4.97 g) was dissolved in CH3CN (ca. 250 mL, dried with 4 A molecular sieves). The solution was stirred under Ar i an ice bath fc 15 min. 2,4,6-trimethyl collidine (1 mL), Hg(CN)2 (0.97 g), HgBr2 (2.67 g), and Hgl2 (0.45 g) WL added to the above solution sequentially. The reaction mixture was then stirred at room temperature for 20 hrs and poured over a suspension of ethyl acetate (200mL) over saturated sodium bicarbonate solution (150 mL). The layers were then separated, and the organic layer was washed with additional aqueous base (3X150 mL). The aqueous washes were combined and extracted ace more with ethyl acetate (200 mL x 1). The organic layers were combined and concentrated to complete dryness to give a yellow colored amorphous solid. A portion of the solid (630 mg) was then dissolved in
a mixmre of CjH5OH and CH3CN (3 mL; 1 : 1, v : v) and loaded onto a gel permeation column (LH-20, 2.8 cm x 90 cm, packed in CH5OH/CH3CN, 1 : 1 v:v).
The column was then developed with HsOH/CH^N (50:50) in which the crude product was dissolved. A total of 11 fractions (ca. 75 ml each) were collected. The sugar-containing fractions (frac. 2-6) indicated by NMR analysis were combined and the solvents were removed by vacuum to give library #2, a light yellow colored solid (376.8 mg, ca. 60% of the crude mixture). The HPLC gel permeation analysis of the crude mixture indicated that the percentage of the oligomerization product with molecular weight about 400 increased from 14% to 27%. The biological activity profiles of the library #2 is shown in Table 1.
Example III.
Conversion of the saccharide Library #2 to a library containing aminoglycosides
(library #3).
In a jacketed hydrogenation tube, ca. 180 mg of library #2 was dissolved in CH3OH (3 mL) with a suspension of palladium hydroxide on carbon (270 mg). A positive pressure of H2 was added (approx 5 to 7 psi) while the tube was mixed vigorously. The reduction process was carried out for 16 hr at ambient temperature. The resultant slurry was then filtered, and the catalyst was sequentially washed with THF, CH3OH, and water. All of the filtrates were combined and concentrated to dryness to give a colorless glass-like solid. A reverse phase thin layer chromatography analysis using C18 (developed with 25%
MeOH in H2O) indicated d e presence of NHX ( NHX = the primary amino NH2 and secondary amino NH, hereafter) by the color development of ninhydrin spray. The solid was then partitioned in ethyl acetate and H2O (50 nJL 1:1, v:v) and die organic layer was separated from the aqueous layer. Both layers were concentrated to dryness. The aqueous layer gave a glassy oil slurry (library #3a, 59.5 mg), whereas the organic layer gave a white colored solid (library #3b, 85.7 mg). A portion (10 uL) of -lie aqueous layer was taken, and spotted onto a silica gel plate (without developing with solvent), spraying with 1% ninhydrin (in CH3OH with trace amount of acetic acid) indicated the presence of NHx functions. The organic portion, which only had a very faint ninhydrin-color-analysis, was analyzed by gel permeation. The biological activities of both library #3a and #3b are shown in Table 1.
Example IV.
Preparation of a saccharide library based on a single carbohydrate (library #4). Raffinose pentahydrate (500mg 0.84 mM) was dissolved ic ^ mL of freshly distilled (over CaH^ pyridine and cooled with an ice bath (5^-10° C) under Ar. Benzylchloroformate (0.4 mL) was then added dropwise to the solution while the solution was vigorously stirred under Ar at 0° to 5° C (ice/ water slurry-bath temperature). The reaction was then continued witii stirring under the above conditions for about 30 min. , at which time methanesulfonyl chloride (0.4 mL) and another portion of benzylchloroformate were added. The reaction mixture was then stirred under the above conditions for another 30 min. followed with the addition of an additional portion of methanesulfonyl chloride (0.4 mL). The entire
reaction mixmre was gradually warmed up to ambient temperature over 45 mins. and then allowed to stir under Ar for another 16 hrs. The pyridinium salt precipitate was then removed by filtration (widi a double-end filter), and die filtrate was directly transferred into a flask containing NaN3 (600 mg). The reaction mixmre was men stirred at room temperature under Ar for 72 hr. The solvent was removed by vacuum (freeze dried over night); the resultant solid was partitioned between 100 mL of 10% MeOH in ethyl acetate (v:v) and water (eq. vol). The organic layer was then washed with HC1 (0.1% cone. HC1 in water; v:v; X2), saturated NaHCO3 (eq. vol; X3) and saturated NaCl (eq. vol.; XI). The crude product was men concentrated to complete dryness to give an oily solid (library #4; 1.41 grams). The biological activity of library #4 is given in Table 1.
Example V .
Conversion of the saccharide library #4 to a library containing aminoglycosides (library #5).
A portion of Library #4 (ca. 700 mg) was suspended in a mixture of organic solvents (ethylacetate and memanol, 1:1, v;v; 50 mL), to which a catalytic amount of palladium oxide on carbon (50 mg) was added. The solution was then purged with hydrogen for 4 hrs with vigorous stirring at ambient temperature. The reaction vessel was sealed under a positive pressure of hydrogen (2-5 psi) and stirred at ambient temperature overnight. The reaction slurry was then filtered, and the catalyst washed with ethyl acetate, memanol and water. All filtrates and washes were combined and concentrated to dryness to give a white amorphous solid. The solid was then partitioned between ethyl acetate (20 mL) and water (20
mL), and the layers were separated from each other with a separatory funnel. The layers were concentrated to dryness. A ninhydrin color test indicated that both fractions contained NHx's. The results of the biological assays of me aqueous fraction, library #5a, and organic fraction, library #5b, are given in Table 1.
Example VI.
Construction of a saccharide library #6 from a single carbohydrate.
Raffinose pentahydrate (2 grams), NaH, (2 grams) and a trace amount of tetrabutylammonium iodide (50 mg) were added to DMF (dried over 4 A molecular sieve), while the reaction mixmre was stirred under Ar over an ice bath. CH3I (430 uL; ca. 2 meq) was added dropwise to the reaction mixmre. The mixmre was then stirred under the above conditions for 35 min followed with an addition of benzylbromide (1.62 mL, 4 meq) dropwise. The reaction temperature was gradually raised to room temperature over 35 to 45 mins. The mixmre was then stirred at the ambient temperature for another 20 hrs before lyophilization (another 48 hrs). The resultant oily solid (light tan colored) was suspended in a methanol/water (50/50, v:v) solution, and freeze-dried again to force a complete removal of the solvent. The dried solid was suspended in denatured etiianol (100 mL) and the insoluble residue was filtered to give a clear filtrate. The filtrate was then concentrated to dryness (9.0 grams). Analysis by reverse phase thin layer chromatography and NMR spectroscopy indicated the formation of multiple sugar- containing products. The crude mixmre was then loaded onto a Toyopearl 40F gel permeation column and eluted with 5% (v.v) methanol in water. Sugar positive fractions were then combined to give a light yellow colored oily mixture of sugars
( library #6; 3.45 gram, 38% of the crude). NMR analysis indicated the existence of multiple OMe (singlets from 2.50 to 3.20 ppm) attached to the sugar moiety. The results of the biological assays of library #6 are given in Table 1.
Example VII.
Conversion of the saccharide library #6 to a library #7 containing aminosugars.
An aliquot of library #6 (3 grams, crude) was dissolved in 50 mL of freshly distilled pyridine (50 mL) and cooled to 0°-5° C with an ice bath. Methanesulfonyl chloride (ca. 1 inL) was then added dropwise for about 80 min. The reaction temperamre was then gradually increased to room temperamre (over 35 to 45 mins.) and d e mixmre was then stirred under Ar for an additional 3 hrs. The resultant slurry was filtered with a double-end filter under reduced pressure with a positive flow of Ar. The filtrate was directly added to a mixture of NaN3 (2.6 gram) and crown emer (catalytic amount 600 mg). The solution was then stirred vigorously under Ar at ambient temperature for 16 hrs. The reaction temperamre was brought to 50° C with an oil-bath under Ar for 2 additional hrs, and cooled to die ambient temperature before filtration. The filtrate was then freeze-dried to remove me pyridine. The resulting oily solid was suspended in a mixmre of solvents (H2O, 3 mL; MeOH, 20 mL; and EtOAc, 10 mL). The milky suspension was loaded onto Toyopearl 40F column which was men eluted wid EtOAc (100%). The fractions containing the OBz (broad multiplets at 7.1 to 7.5 ppm. C6H5) and OMe (multiplets at 2.6 to 2.8 ppm) signals by proton NMR analysis were combined and concentrated to dryness (library #7a, 450 mg). The NMR analysis also indicated the existence of the CH-N3 functional groups by the
broad multiplets at the 2.8 to 3.4 ppm. The biological testing result of library #7a is shown in Table 1.
Library #7a was then suspended in a mixmre of solvents (20 mL, H3OH/EtOAc, 50:50, v:v) in the presence of a catalytic amount of Palladium oxide on carbon (50 mg). The reaction vessel was then purged widi a positive flow of hydrogen with vigorous stirring for 5 hrs. The reaction vessel was then sealed while connected to a hydrogen tank with a hydrogen pressure at 2.5 psi. and the pressure was gradually raised to 15 psi during the next 72 hrs. The reaction mixture was then filtered, and die catalyst was washed sequentially witi H2O, MeOH and EtOAc. All washes and filtrates were combined and concentrated to dryness to give a pale white solid. The solid was tiien partitioned between 250 mL of water and 250 mL of EtOAc. The layers was then separated. The organic layer was washed once again with water, while the aqueous layer was washed once widi EtOAc. The washes were then combined widi the corresponding layers of extracts and solutions were then concentrated to dryness. The aqueous fraction gave a clear oily solid Gibrary #7b, 194 mg) whereas the organic layer gave a light yellow colored solid (library #7c, 176 mg). The biological activity test results of the libraries #7b and 7c are given in Table 1.
Table 1. Antimicrobial Profiles of different saccharide libraries
Claims
1. A method for generating and screening a library of compounds, comprising: reacting one or more polyhydroxyl-containing organic compounds with subequimolar amounts of displacing groups to form a first mixture comprising displacing group-substituted polyhydroxyl-containing organic compounds; testing said first mixture or a reaction product made tiierefrom for the presence of a component having a biological activity.
2. The method of claim 1 further comprising the step of: oligomerizing a portion of said first mixmre to form a second mixmre comprising displacing group-substituted polyhydroxyl-containing organic oligomers.
3. The method of claim 1 further comprising the step of: reducing a portion of said first mixture comprising displacing group- substituted polyhydroxyl-containing organic compounds to form a second mixmre comprising displacing group-substituted, reduced polyhydroxyl-containing organic compounds.
4. The method of claim 1 further comprising the steps of: oligomerizing a portion of said first mixture to form a second mixture comprising displacing group-substituted polyhydroxyl-containing organic oligomers; reducing a portion of said second mixmre comprising displacing group-substituted polyhydroxyl-containing organic oligomers to form a third mixture comprising displacing group-substituted, reduced polyhydroxyl-containing organic oligomers.
5. The method of claim 1 further comprising the step of: alkylating a portion of said first mixture.
6. The method of claim 2 further comprising the step of: alkylating a portion of said first or second mixtures.
7. The method of claim 3 further comprising the step of: alkylating a portion of said first or second mixtures.
8. The method of claim 4 further comprising the step of: alkylating a portion of said first or second or third mixtures.
9. The method of claim 1 further comprising the step of: halogenating a portion of said first mixmre.
10. The method of claim 2 further comprising the step of: halogenating a portion of said first or second mixtures.
11. The method of claim 3 further comprising the step of: halogenating a portion of said first or second mixtures.
12. The method of claim 4 further comprising the step of: halogenating a portion of said first or second or third mixtures.
13. The method of claim 1 wherein hydroxyl groups on said polyhydroxyl-containing organic compounds are activated prior to reaction with displacing groups to form organic compounds containing activated hydroxyl functionalities.
14. The method of claim 13 wherein said activation is accomplished by reaction of said polyhydroxyl-containing organic compounds widi a bromine or iodine salt.
15. The method of claim 13 wherein said activation is accomplished by reaction of said polyhydroxyl-containing organic compounds widi methanesulfonyl chloride.
16. The method of claim 13 wherein said activation is accomplished by reaction of said polyhydroxyl-containing organic compounds wid alkylsulfonyl chloride.
17. The method of claim 13 wherein said activated hydroxyl functionalities of said compounds are sulfonyl esters.
18. The method of claim 2 wherein said hydroxyl groups on said polyhydroxyl-containing organic compounds are activated prior to said step of oligomerizing.
19. The method of claim 18 wherein said activation is accomplished by reacting said polyhydroxyl-containing organic compounds with a sulfonyl compound.
20. The memod of claim 18 wherein said activation is accomplished by reacting said polyhydroxyl-containing organic compounds with a halide compound.
21. The memod of claim 18 wherein said activation is accomplished by reacting said polyhydroxyl-containing organic compounds with a thiolate compound.
22. The method of claim 13 or 18 wherein the amount of activated hydroxyl functionalities formed is controlled by first reacting said polyhydroxyl- containing organic compounds widi subequimolar amounts of protecting groups.
23. The method of claim 1 further comprising the steps of: fractionating a mixture which demonstrates biological activity to form fractions; testing said fractions for said biological activity.
24. The method of claim 23 wherein said steps of fractionating and testing are repeated until an active ingredient has been purified.
25. The method of claim 1 wherein the displacing group is a carbanion.
26. The method of claim 1 wherein the displacing group is a halogen ion.
27. The method of claim 1 wherein die displacing group is an azide ion.
28. The method of claim 1 wherein me displacing group is a nitrile.
29. The method of claim 1 wherein the displacing group is a sulfite.
30. The method of claim 1 wherein the displacing group is a tiiiolate.
31. The method of claim 1 wherein the displacing group is an alkoxide.
32. The method of claim 1 wherein a mixture of displacing groups is used in said step of reacting.
33. A method for generating and screening a library of compounds, comprising: reacting one or more polyhydroxyl-containing organic compounds with subequimolar amounts of protecting groups to form a first mixture of randomly protected polyhydroxyl-containing organic compounds; oxidizing said first mixmre to form a second mixture comprising carbonyl-containing organic compounds; testing said first or said second mixtures or a reaction product made therefrom for die presence of a component having a biological activity.
34. The method of claim 33 wherein said carbonyl-containing organic compounds are reacted with carbanions to form branched-chain, organic oligomers.
35. The method of claim 33 wherein said carbonyl-containing organic compounds are reacted with a Wittig reagent.
36. The method of claim 1 further comprising the step of: oxidizing a portion of said first mixmre to form a second mixmre comprising carbonyl-containing organic compounds.
37. The metiiod of claim 2 further comprising the step of: oxidizing a portion of said first or second mixmre to form a tiiird mixture comprising carbonyl-containing organic compounds.
38. The metiiod of claim 3 further comprising d e step of: oxidizing a portion of said first or second mixmre to form a third mixture comprising carbonyl-containing organic compounds.
39. The method of claim 4 further comprising the step of: oxidizing a portion of said first or second or tiiird mixture to form a fourth mixture comprising carbonyl-containing organic compounds.
40. The method of any of claims 36-39 further comprising the step of: reacting one or more polyhydroxyl-containing organic compounds with subequimolar amounts of protecting groups to form a mixture of randomly protected polyhydroxyl-containing organic compounds, prior to the step of oxidizing.
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PCT/US1995/001389 WO1995021850A1 (en) | 1994-02-09 | 1995-02-08 | Generation and screening of synthetic drug libraries |
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EP (1) | EP0743952A1 (en) |
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Families Citing this family (268)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6030776A (en) * | 1990-06-11 | 2000-02-29 | Nexstar Pharmaceuticals, Inc. | Parallel SELEX |
RU2134693C1 (en) * | 1993-02-23 | 1999-08-20 | Зе Трастис оф Принстон Юниверсити | Methods of forming glycoside bonds, chemical composition, glycoside and glycoside library |
US20030099945A1 (en) * | 1994-09-20 | 2003-05-29 | Invenux, Inc. | Parallel selex |
US6048698A (en) * | 1994-09-20 | 2000-04-11 | Nexstar Pharmaceuticals, Inc. | Parallel SELEX™ |
DE19642751A1 (en) * | 1996-10-16 | 1998-04-23 | Deutsches Krebsforsch | Saccharide library |
US5965719A (en) * | 1996-11-15 | 1999-10-12 | Sunsorb Biotech, Inc. | Combinatorial synthesis of carbohydrate libraries |
US7252950B1 (en) * | 1997-09-04 | 2007-08-07 | The Regents Of The University Of California | Assays for detecting modulators of cytoskeletal function |
US6960457B1 (en) * | 1997-09-04 | 2005-11-01 | Stanford University | Reversible immobilization of arginine-tagged moieties on a silicate surface |
CA2323725A1 (en) | 1998-03-20 | 1999-09-23 | The Rockefeller University | Assays for screening compounds which interact with cation channel proteins, mutant prokaryotic cation channel proteins, and uses thereof |
US6316616B1 (en) | 1998-04-02 | 2001-11-13 | President And Fellows Of Harvard College | Parallel combinatorial approach to the discovery and optimization of catalysts and uses thereof |
JP2002522747A (en) | 1998-04-14 | 2002-07-23 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Assays for detecting microtubule depolymerization inhibitors |
US6872537B1 (en) | 1998-04-14 | 2005-03-29 | Regents Of The University Of California | Assays for the detection of microtubule depolymerization inhibitors |
US6699969B1 (en) | 1998-04-14 | 2004-03-02 | The Regents Of The University Of California | Assays for the detection of microtubule depolymerization inhibitors |
US7402400B2 (en) | 2001-07-03 | 2008-07-22 | Regents Of The University Of California | Mammalian sweet taste receptors |
US6548733B2 (en) | 1998-12-21 | 2003-04-15 | The Genetics Company, Inc. | Function-based small molecular weight compound screening system in drosophila melanogaster |
US6649358B1 (en) | 1999-06-01 | 2003-11-18 | Caliper Technologies Corp. | Microscale assays and microfluidic devices for transporter, gradient induced, and binding activities |
EP1230344B1 (en) | 1999-11-17 | 2011-11-02 | Mendel Biotechnology, Inc. | Plant biochemistry-related genes |
EP1950306A1 (en) | 1999-11-17 | 2008-07-30 | Mendel Biotechnology, Inc. | Environmental stress tolerance genes |
US6458538B1 (en) | 1999-12-14 | 2002-10-01 | Ptc Therapeutics, Inc. | Methods of assaying for compounds that inhibit premature translation termination and nonsense-mediated RNA decay |
US8642284B1 (en) | 1999-12-15 | 2014-02-04 | Massachusetts Institute Of Technology | Methods for identifying agents that alter NAD-dependent deacetylation activity of a SIR2 protein |
US7452664B2 (en) * | 1999-12-15 | 2008-11-18 | Massachusetts Institute Of Technology | Methods for identifying agents which alter histone protein acetylation |
AU2000234813A1 (en) | 2000-02-03 | 2001-08-14 | The Regents Of The University Of California | Assays for the detection of agents that affect cognition |
TW201006846A (en) | 2000-03-07 | 2010-02-16 | Senomyx Inc | T1R taste receptor and genes encidung same |
WO2002015675A1 (en) | 2000-08-22 | 2002-02-28 | Mendel Biotechnology, Inc. | Genes for modifying plant traits iv |
US7045283B2 (en) | 2000-10-18 | 2006-05-16 | The Regents Of The University Of California | Methods of high-throughput screening for internalizing antibodies |
US7572575B2 (en) * | 2000-12-13 | 2009-08-11 | Massachusetts Institute Of Technology | SIR2 activity |
TW201022287A (en) | 2001-01-03 | 2010-06-16 | Senomyx Inc | T1R taste receptors and genes encoding same |
US7883856B2 (en) | 2001-04-05 | 2011-02-08 | Senomyx Inc. | Identification of bitter ligands that specifically activate human T2R receptors and related assays for identifying human bitter taste modulators |
US20030191073A1 (en) | 2001-11-07 | 2003-10-09 | Challita-Eid Pia M. | Nucleic acid and corresponding protein entitled 161P2F10B useful in treatment and detection of cancer |
US7803982B2 (en) | 2001-04-20 | 2010-09-28 | The Mount Sinai School Of Medicine Of New York University | T1R3 transgenic animals, cells and related methods |
EP1572871A4 (en) * | 2001-04-20 | 2007-11-14 | Sinai School Medicine | T1r3 a novel taste receptor |
WO2003001876A2 (en) | 2001-06-26 | 2003-01-09 | Senomyx, Inc. | T1r hetero-oligomeric taste receptors and cell lines that express said receptors and use thereof for identification of taste compounds |
WO2003031604A1 (en) * | 2001-10-12 | 2003-04-17 | The Regents Of The University Of California | Gastrointestinal chemosensory receptors |
US7871619B2 (en) | 2001-11-30 | 2011-01-18 | Chemocentryx, Inc. | Compositions and methods for detecting and treating diseases and conditions related to chemokine receptors |
WO2003048341A2 (en) * | 2001-12-03 | 2003-06-12 | Uab Research Foundation | Cap-gly domain structure and uses thereof |
WO2003051314A2 (en) * | 2001-12-18 | 2003-06-26 | Medallion Biomedical, Llc | Antibiotic compounds |
JP2005523688A (en) * | 2002-01-18 | 2005-08-11 | ブリストル−マイヤーズ スクイブ カンパニー | Identification of polynucleotides and polypeptides for predicting the activity of protein tyrosine kinases and / or compounds that interact with protein tyrosine kinase pathways |
EP1572906A4 (en) * | 2002-03-04 | 2008-05-28 | Bristol Myers Squibb Co | Novel nucleic acid molecules and polypeptides encoding baboon tafi |
US7893218B2 (en) | 2003-06-16 | 2011-02-22 | Stowers Institute For Medical Research | Antibodies that specifically bind SOST peptides |
WO2004016733A2 (en) | 2002-08-16 | 2004-02-26 | Agensys, Inc. | Nucleic acid and corresponding protein entitled 251p5g2 useful in treatment and detection of cancer |
EP3249046B1 (en) | 2002-09-18 | 2020-07-08 | Mendel Biotechnology, Inc. | Polynucleotides and polypeptides in plants |
US20040171037A1 (en) * | 2002-11-19 | 2004-09-02 | Jing Li | Amplified genes involved in cancer |
EP1585815A4 (en) * | 2003-01-21 | 2006-02-22 | Bristol Myers Squibb Co | Polynucleotide encoding a novel acyl coenzyme a, monoacylglycerol acyltransferase-3 (mgat3), and uses thereof |
NZ578591A (en) | 2003-05-30 | 2011-01-28 | Agensys Inc | Prostate stem cell antigen (PSCA) variants and subsequences thereof, namely SEQ ID NO: 6554 |
US20050026194A1 (en) * | 2003-06-20 | 2005-02-03 | Tularik Inc. | Gene amplification and overexpression in cancer |
WO2005002527A2 (en) * | 2003-07-03 | 2005-01-13 | Massachusetts Institute Of Technology | Sirt1 modulation of adipogenesis and adipose function |
NZ619746A (en) | 2003-08-06 | 2014-05-30 | Senomyx Inc | Novel flavors, flavor modifiers, tastants, taste enhancers, umami or sweet tastants, and/or enhancers and use thereof |
US20050244810A1 (en) * | 2003-09-29 | 2005-11-03 | Egan Josephine M | Taste signaling in gastrointestinal cells |
US7480382B2 (en) * | 2003-09-30 | 2009-01-20 | Microsoft Corporation | Image file container |
JP4796967B2 (en) | 2003-11-07 | 2011-10-19 | ヴァーミリオン インコーポレイテッド | Biomarkers for Alzheimer's disease |
US20050164969A1 (en) * | 2003-11-19 | 2005-07-28 | Massachusetts Institute Of Technology | Method of extending life span |
AU2004304665B2 (en) | 2003-12-24 | 2009-03-12 | Novo Nordisk A/S | Transgenic non-human mammal comprising a polynucleotide encoding human or humanized C5aR |
EP1722771B1 (en) | 2004-03-02 | 2014-04-23 | McGill University | Compositions and methods for preventing or treating an inflammatory response |
JP5223072B2 (en) | 2004-04-02 | 2013-06-26 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | Methods and compositions for treating and preventing diseases associated with avβ5 integrin |
US7794713B2 (en) * | 2004-04-07 | 2010-09-14 | Lpath, Inc. | Compositions and methods for the treatment and prevention of hyperproliferative diseases |
US7939251B2 (en) | 2004-05-06 | 2011-05-10 | Roche Molecular Systems, Inc. | SENP1 as a marker for cancer |
US7323347B2 (en) * | 2004-05-27 | 2008-01-29 | Sensata Technologies, Inc. | Biosensor surface structures and methods |
NZ551627A (en) | 2004-05-28 | 2010-02-26 | Agensys Inc | Antibodies and related molecules that bind to PSCA proteins |
EP1766077B1 (en) | 2004-06-21 | 2012-03-28 | The Board Of Trustees Of The Leland Stanford Junior University | Genes and pathways differentially expressed in bipolar disorder and/or major depressive disorder |
WO2006023410A2 (en) | 2004-08-20 | 2006-03-02 | Buck Institute For Age Research | Small molecules that replace or agonize p53 function |
US7850960B2 (en) | 2004-12-30 | 2010-12-14 | University Of Washington | Methods for regulation of stem cells |
WO2006094014A2 (en) | 2005-02-28 | 2006-09-08 | The Regents Of The University Of California | Methods for diagnosis and treatment of endometrial cancer |
EP2444099A1 (en) | 2005-03-31 | 2012-04-25 | Agensys, Inc. | Antibodies and related molecules that bind to 161P2F10B proteins |
US8318906B2 (en) | 2005-04-15 | 2012-11-27 | The Regents Of The University Of California | EMP2 antibodies and their therapeutic uses |
US20070099830A1 (en) | 2005-04-21 | 2007-05-03 | Massachusetts Institute Of Technology | Sirt4 activities |
EP2392258B1 (en) | 2005-04-28 | 2014-10-08 | Proteus Digital Health, Inc. | Pharma-informatics system |
DK3467096T3 (en) | 2005-06-07 | 2022-03-28 | Dsm Nutritional Products Ag | EUKARYOTE MICRO-ORGANISMS FOR THE PRODUCTION OF LIPIDS AND ANTIOXIDANTS |
PL1904093T3 (en) | 2005-07-19 | 2018-12-31 | Stemgen S.P.A. | Inhibition of the tumorigenic potential of tumor stem cells by bmp-4 |
WO2007016597A2 (en) * | 2005-07-29 | 2007-02-08 | The Regents Of The University Of California | Targeting tnf-alpha converting enzyme (tace)-dependent growth factor shedding in cancer therapy |
US20070072247A1 (en) * | 2005-08-31 | 2007-03-29 | Academia Sinica | Methods and reagents for the analysis and purification of polysaccharides |
US7943134B2 (en) * | 2005-08-31 | 2011-05-17 | Academia Sinica | Compositions and methods for identifying response targets and treating flavivirus infection responses |
US7781209B2 (en) * | 2005-09-25 | 2010-08-24 | Multispan, Inc. | GPCR-expressing cell lines |
CA2623705A1 (en) * | 2005-09-27 | 2007-04-05 | Bernd Helmut Adam Rehm | Polymer particles and uses thereof |
JP5342878B2 (en) | 2005-10-20 | 2013-11-13 | セノミクス・インコーポレーテッド | Chimeric human sweetness-umami and umami-sweet taste receptors |
US20080213274A1 (en) * | 2005-10-28 | 2008-09-04 | Sabbadini Roger A | Compositions and methods for the treatment and prevention of fibrotic, inflammatory, and neovascularization conditions of the eye |
US20090074720A1 (en) * | 2005-10-28 | 2009-03-19 | Sabbadini Roger A | Methods for decreasing immune response and treating immune conditions |
US7749730B2 (en) * | 2005-11-03 | 2010-07-06 | Redpoint Bio Corporation | High throughput screening assay for the TRPM5 ion channel |
CA2628238A1 (en) | 2005-11-07 | 2007-05-18 | The Scripps Research Institute | Compositions and methods for controlling tissue factor signaling specificity |
EP2564864B8 (en) | 2005-11-12 | 2015-05-13 | The Board of Trustees of The Leland Stanford Junior University | FGF2-related methods for diagnosing and treating depression |
EP1976567B1 (en) | 2005-12-28 | 2020-05-13 | The Scripps Research Institute | Natural antisense and non-coding rna transcripts as drug targets |
GB0605584D0 (en) * | 2006-03-20 | 2006-04-26 | Olink Ab | Method for analyte detection using proximity probes |
US20100278821A1 (en) | 2006-03-21 | 2010-11-04 | The Regents Of The University Of California | N-cadherin: target for cancer diagnosis and therapy |
CA2646597A1 (en) | 2006-03-21 | 2007-09-27 | The Regents Of The University Of California | N-cadherin and ly6 e: targets for cancer diagnosis and therapy |
US20070232556A1 (en) * | 2006-03-31 | 2007-10-04 | Montine Thomas J | Methods and compositions for the treatment of neurological diseases and disorders |
ES2640453T3 (en) | 2006-04-21 | 2017-11-03 | Senomyx, Inc. | Processes for preparing solid flavoring compositions |
CN101421419B (en) | 2006-05-02 | 2012-08-29 | 学校法人帝京大学 | Method for screening of substance capable of increasing glutathione |
US7862812B2 (en) * | 2006-05-31 | 2011-01-04 | Lpath, Inc. | Methods for decreasing immune response and treating immune conditions |
US20100240029A1 (en) * | 2006-06-06 | 2010-09-23 | Massachusetts Institute Of Technology | Cholesterol-Regulating Complex of SIRT1 and LXR and Methods of Use |
DE602006012460D1 (en) | 2006-06-07 | 2010-04-08 | Nutrinova Gmbh | Method for screening substances that modulate the activity of Gpcrs |
US7572618B2 (en) | 2006-06-30 | 2009-08-11 | Bristol-Myers Squibb Company | Polynucleotides encoding novel PCSK9 variants |
US7674594B2 (en) * | 2006-07-27 | 2010-03-09 | Redpoint Bio Corporation | Screening assay for inhibitors of TRPA1 activation by a lower alkyl phenol |
MX2009001346A (en) * | 2006-08-01 | 2009-04-17 | Ocean Nutrition Canada Ltd | Oil producing microbes and methods of modification thereof. |
WO2008022035A2 (en) * | 2006-08-10 | 2008-02-21 | The Scripps Research Institute | Methods for identifying cellular modulators of disaggregation activity or aggregation activity in an animal |
US20100316626A1 (en) * | 2006-08-11 | 2010-12-16 | The Government Of The United States Of America As Represented By The Secretary | Methods for treatment and diagnosis of psychiatric disorders |
EP2061900A2 (en) | 2006-08-25 | 2009-05-27 | Oncotherapy Science, Inc. | Prognostic markers and therapeutic targets for lung cancer |
EP2062049B1 (en) | 2006-09-06 | 2014-04-30 | The Regents of the University of California | Molecular diagnosis and classification of malignant melanoma |
US20100062000A1 (en) * | 2006-11-21 | 2010-03-11 | The Regents Of The University Of California | Rhamm, a Co-Receptor and Its Interactions with Other Receptors in Cancer Cell Motility and the Identification of Cancer Prognitor Cell Populations |
US20080187928A1 (en) * | 2006-12-29 | 2008-08-07 | The Salk Institute For Biological Studies | Methods for enhancing exercise performance |
US20080229436A1 (en) * | 2007-03-15 | 2008-09-18 | Board Of Trustees Of The University Of Arkansas | Methods for identifying modulators of lifespan and resistance to oxidative stress |
ES2477292T3 (en) | 2007-08-13 | 2014-07-16 | Baxter International Inc. | IVIG modulation of chemokines for the treatment of multiple sclerosis, Alzheimer's disease and Parkinson's disease |
CA2982520A1 (en) | 2007-08-21 | 2009-02-26 | Senomyx, Inc. | Identification of human t2r receptors that respond to bitter compounds that elicit the bitter taste in compositions, and the use thereof in assays to identify compounds that inhibit (block) bitter taste in compositions and use thereof |
FI2192946T3 (en) | 2007-09-25 | 2022-11-30 | In-body device with virtual dipole signal amplification | |
CA2702025A1 (en) * | 2007-10-01 | 2009-04-09 | Redpoint Bio Corporation | A non-desensitizing mutant of the transient receptor potential trpm5 ion channel |
EP3241846B1 (en) | 2007-10-04 | 2022-02-23 | ZymoGenetics, Inc. | B7 family member zb7h6 and related compositions and methods |
CA2703165A1 (en) | 2007-10-22 | 2009-04-30 | The Regents Of The University Of California | Biomarkers for prenatal diagnosis of congenital cytomegalovirus |
CA2703519C (en) | 2007-11-09 | 2017-04-18 | The Salk Institute For Biological Studies | Use of tam receptor inhibitors as immunoenhancers and tam activators as immunosuppressors |
US20090203605A1 (en) * | 2008-02-01 | 2009-08-13 | The Scripps Research Institute | Methods For Treating A Condition Characterized By Dysfunction In Protein Homeostasis |
ES2489840T3 (en) | 2008-03-05 | 2014-09-02 | The Regents Of The University Of California | Molecular prognosis and classification of malignant melanoma based on markers selected from the list consisting of RGS1, NCOA3, SPP1, PHIP |
MX2010010165A (en) | 2008-03-17 | 2010-11-25 | Scripps Research Inst | Combined chemical and genetic approaches for generation of induced pluripotent stem cells. |
CN102083850B (en) | 2008-04-21 | 2015-08-12 | 加利福尼亚大学董事会 | Selectivity high-affinity polydentate ligand and preparation method thereof |
JP2011529708A (en) | 2008-08-04 | 2011-12-15 | ユニバーシティ オブ マイアミ | Innate immune response regulator STING (interferon gene stimulator) |
CA2738019A1 (en) | 2008-09-23 | 2010-04-08 | President And Fellows Of Harvard College | Sirt4 and uses thereof |
US8153606B2 (en) * | 2008-10-03 | 2012-04-10 | Opko Curna, Llc | Treatment of apolipoprotein-A1 related diseases by inhibition of natural antisense transcript to apolipoprotein-A1 |
JP5798037B2 (en) | 2008-11-06 | 2015-10-21 | ユニバーシティ オブ マイアミ | Role of soluble uPAR in the pathogenesis of proteinuria |
EP2352847B1 (en) | 2008-11-10 | 2014-01-08 | The United States of America, as represented by The Secretary, Department of Health and Human Services | Gene signature for predicting prognosis of patients with solid tumors |
EP3623374B1 (en) | 2008-12-03 | 2021-09-08 | The Scripps Research Institute | Stem cell cultures |
WO2010065671A2 (en) | 2008-12-04 | 2010-06-10 | Curna, Inc. | Treatment of vascular endothelial growth factor (vegf) related diseases by inhibition of natural antisense transcript to vegf |
CN102317458B (en) * | 2008-12-04 | 2018-01-02 | 库尔纳公司 | Pass through treatment of the suppression of erythropoietin(EPO) (EPO) natural antisense transcript to EPO relevant diseases |
RU2746478C2 (en) | 2008-12-04 | 2021-04-14 | КьюРНА, Инк. | Treatment of tumors of diseases related to the genom-suppressor by therapy of natural transcript inhibition in anti-significant orientation regarding this gene |
EP2370175A2 (en) | 2008-12-16 | 2011-10-05 | Bristol-Myers Squibb Company | Methods of inhibiting quiescent tumor proliferation |
EP3312269A1 (en) | 2008-12-17 | 2018-04-25 | The Scripps Research Institute | Generation and maintenance of stem cells |
PT2396038E (en) | 2009-02-12 | 2016-02-19 | Curna Inc | Treatment of brain derived neurotrophic factor (bdnf) related diseases by inhibition of natural antisense transcript to bdnf |
ES2656290T3 (en) | 2009-03-16 | 2018-02-26 | Curna, Inc. | Treatment of diseases related to nuclear factor (derived from erythroid 2) similar to 2 (NRF2) by inhibition of natural antisense transcript to NRF2 |
EP2408920B1 (en) | 2009-03-17 | 2017-03-08 | CuRNA, Inc. | Treatment of delta-like 1 homolog (dlk1) related diseases by inhibition of natural antisense transcript to dlk1 |
US20100240069A1 (en) * | 2009-03-18 | 2010-09-23 | Boehringer Ingelheim International Gmbh | Drug Discovery Assay for Modulators of HIF-Prolyl Hydroxylase Activity |
US20100240065A1 (en) * | 2009-03-18 | 2010-09-23 | Boehringer Ingelheim International Gmbh | Prolyl Hydroxylase Compositions and Methods of Use Thereof |
CA2756760A1 (en) * | 2009-03-27 | 2010-11-04 | Gojo Industries, Inc. | Compositions and methods for screening and using compounds antagonizing spore-surface interactions |
MX2011011506A (en) | 2009-04-28 | 2012-05-08 | Proteus Biomedical Inc | Highly reliable ingestible event markers and methods for using the same. |
KR101722541B1 (en) | 2009-05-06 | 2017-04-04 | 큐알엔에이, 인크. | Treatment of tristetraproline(ttp) related diseases by inhibition of natural antisense transcript to ttp |
CN103223177B (en) | 2009-05-06 | 2016-08-10 | 库尔纳公司 | By suppression therapy lipid transfer and the metabolic gene relevant disease of the natural antisense transcript for lipid transfer and metabolic gene |
US9012139B2 (en) | 2009-05-08 | 2015-04-21 | Curna, Inc. | Treatment of dystrophin family related diseases by inhibition of natural antisense transcript to DMD family |
CA2762369C (en) | 2009-05-18 | 2021-12-28 | Joseph Collard | Treatment of reprogramming factor related diseases by inhibition of natural antisense transcript to a reprogramming factor |
EP2432882B1 (en) | 2009-05-22 | 2019-12-25 | CuRNA, Inc. | TREATMENT OF TRANSCRIPTION FACTOR E3 (TFE3) and INSULIN RECEPTOR SUBSTRATE 2 (IRS2) RELATED DISEASES BY INHIBITION OF NATURAL ANTISENSE TRANSCRIPT TO TFE3 |
CA2764683A1 (en) | 2009-05-28 | 2010-12-02 | Joseph Collard | Treatment of antiviral gene related diseases by inhibition of natural antisense transcript to an antiviral gene |
JP6128846B2 (en) | 2009-06-16 | 2017-05-17 | クルナ・インコーポレーテッド | Treatment of PON1 gene-related diseases by suppression of natural antisense transcripts against paraoxonase (PON1) |
EP2443237B1 (en) | 2009-06-16 | 2017-02-22 | CuRNA, Inc. | Treatment of collagen gene related diseases by inhibition of natural antisense transcript to a collagen gene |
CN102597238B (en) | 2009-06-24 | 2016-06-29 | 库尔纳公司 | The relevant disease of TNFR2 is treated by suppressing for the natural antisense transcript of tumor necrosis factor receptor 2 (TNFR2) |
CN102482672B (en) | 2009-06-26 | 2016-11-09 | 库尔纳公司 | By suppressing the natural antisense transcript treatment Down syndrome gene-associated diseases of Down syndrome gene |
US20110021362A1 (en) * | 2009-07-20 | 2011-01-27 | Constellation Pharmaceuticals | Agents for stimulating activity of methyl modifying enzymes and methods of use thereof |
US10087252B2 (en) | 2009-07-24 | 2018-10-02 | The Regents Of The University Of California | Methods and compositions for treating and preventing disease associated with αvβ5 integrin |
CA2768947C (en) | 2009-07-24 | 2018-06-19 | Opko Curna, Llc | Treatment of sirtuin (sirt) related diseases by inhibition of natural antisense transcript to a sirtuin (sirt) |
CN102762731B (en) | 2009-08-05 | 2018-06-22 | 库尔纳公司 | By inhibiting to treat insulin gene (INS) relevant disease for the natural antisense transcript of insulin gene (INS) |
KR101827015B1 (en) | 2009-08-11 | 2018-02-07 | 큐알엔에이, 인크. | Treatment of adiponectin (adipoq) related diseases by inhibition of natural antisense transcript to an adiponectin (adipoq) |
CN102576022B (en) | 2009-08-14 | 2016-06-15 | 加利福尼亚大学董事会 | The method diagnosing and treating infantile autism |
US20120148604A1 (en) | 2009-08-20 | 2012-06-14 | Transposagen Biopharmaceuticals, Inc. | Trp inhibitors and uses thereof |
EP2982755B1 (en) | 2009-08-21 | 2020-10-07 | CuRNA, Inc. | Treatment of 'c terminus of hsp70-interacting protein' (chip) related diseases by inhibition of natural antisense transcript to chip |
JP5964232B2 (en) | 2009-08-25 | 2016-08-03 | カッパーアールエヌエー,インコーポレイテッド | Treatment of IQGAP-related diseases by inhibition of natural antisense transcripts against 'IQ motif-containing GTPase-activating protein' (IQGAP) |
WO2011038210A2 (en) | 2009-09-25 | 2011-03-31 | Curna, Inc. | Treatment of filaggrin (flg) related diseases by modulation of flg expression and activity |
CA2812929C (en) * | 2009-11-12 | 2015-02-03 | Texas Tech University | Compositions and methods for treating hyperproliferative disorders |
ES2365343B1 (en) | 2009-11-19 | 2012-07-10 | Fundación Centro Nacional De Investigaciones Cardiovasculares Carlos Iii | USE OF CD98 AS AN ENDOMETRIAL RECEPTIVITY MARKER. |
EP3348277A1 (en) | 2009-11-20 | 2018-07-18 | The Regents of The University of California | Epithelial membrane protein-2 (emp2) and proliferative vitreoretinopathy (pvr) |
KR101823702B1 (en) | 2009-12-16 | 2018-01-30 | 큐알엔에이, 인크. | Treatment of membrane bound transcription factor peptidase, site 1 (mbtps1) related diseases by inhibition of natural antisense transcript to mbtps1 |
JP6031356B2 (en) | 2009-12-23 | 2016-11-24 | カッパーアールエヌエー,インコーポレイテッド | Treatment of uncoupling protein 2 (UCP2) -related diseases by inhibition of natural antisense transcripts against UCP2. |
US8940708B2 (en) | 2009-12-23 | 2015-01-27 | Curna, Inc. | Treatment of hepatocyte growth factor (HGF) related diseases by inhibition of natural antisense transcript to HGF |
CN102782134B (en) | 2009-12-29 | 2017-11-24 | 库尔纳公司 | NRF1 relevant diseases are treated by suppressing the natural antisense transcript of the core breathing factor 1 (NRF1) |
CA2785177C (en) | 2009-12-29 | 2019-09-24 | Curna, Inc. | Treatment of tumor protein 63 (p63) related diseases by inhibition of natural antisense transcript to p63 |
JP6083735B2 (en) | 2009-12-31 | 2017-02-22 | カッパーアールエヌエー,インコーポレイテッド | Treatment of insulin receptor substrate 2 (IRS2) related diseases by inhibition of natural antisense transcripts against insulin receptor substrate 2 (IRS2) and transcription factor 3 (TFE3) |
WO2011082409A2 (en) | 2010-01-04 | 2011-07-07 | Curna, Inc. | Treatment of interferon regulatory factor 8 (irf8) related diseases by inhibition of natural antisense transcript to irf8 |
WO2011085066A2 (en) | 2010-01-06 | 2011-07-14 | Curna, Inc. | Treatment of pancreatic developmental gene related diseases by inhibition of natural antisense transcript to a pancreatic developmental gene |
EP2524039B1 (en) | 2010-01-11 | 2017-11-29 | CuRNA, Inc. | Treatment of sex hormone binding globulin (shbg) related diseases by inhibition of natural antisense transcript to shbg |
RU2611192C2 (en) | 2010-01-25 | 2017-02-21 | Курна, Инк. | TREATMENT OF RNase H1 RELATED DISEASES BY INHIBITION OF NATURAL ANTISENSE TRANSCRIPT TO RNase H1 |
WO2011091435A2 (en) | 2010-01-25 | 2011-07-28 | Mount Sinai School Of Medicine | Methods of treating liver disease |
WO2011094759A2 (en) | 2010-02-01 | 2011-08-04 | The Regents Of The University Of California | Novel diagnostic and therapeutic targets associated with or regulated by n-cadherin expression and/or epithelial to mesenchymal transition (emt) in prostate cancer and other malignancies |
US8962586B2 (en) | 2010-02-22 | 2015-02-24 | Curna, Inc. | Treatment of pyrroline-5-carboxylate reductase 1 (PYCR1) related diseases by inhibition of natural antisense transcript to PYCR1 |
GB201004292D0 (en) * | 2010-03-15 | 2010-04-28 | Olink Ab | Assay for localised detection of analytes |
CA2795145C (en) | 2010-04-02 | 2019-01-22 | Curna, Inc. | Treatment of colony-stimulating factor 3 (csf3) related diseases by inhibition of natural antisense transcript to csf3 |
TWI644675B (en) | 2010-04-09 | 2018-12-21 | 可娜公司 | Treatment of fibroblast growth factor 21 (fgf21) related diseases by inhibition of natural antisense transcript to fgf21 |
KR101915115B1 (en) | 2010-05-03 | 2018-11-05 | 큐알엔에이, 인크. | Treatment of sirtuin (sirt) related diseases by inhibition of natural antisense transcript to a sirtuin (sirt) |
TWI531370B (en) | 2010-05-14 | 2016-05-01 | 可娜公司 | Treatment of par4 related diseases by inhibition of natural antisense transcript to par4 |
US8895528B2 (en) | 2010-05-26 | 2014-11-25 | Curna, Inc. | Treatment of atonal homolog 1 (ATOH1) related diseases by inhibition of natural antisense transcript to ATOH1 |
CN102971423B (en) | 2010-05-26 | 2018-01-26 | 库尔纳公司 | MSRA relevant diseases are treated by suppressing Methionine Sulfoxide Reductase A (MSRA) natural antisense transcript |
EP2400304A1 (en) | 2010-06-22 | 2011-12-28 | Centro de Investigación Cooperativa En Biomateriales ( CIC biomaGUNE) | Method for the characterization of intermolecular interactions |
PT2585596T (en) | 2010-06-23 | 2021-03-23 | Curna Inc | Treatment of sodium channel, voltage-gated, alpha subunit (scna) related diseases by inhibition of natural antisense transcript to scna |
US8980860B2 (en) | 2010-07-14 | 2015-03-17 | Curna, Inc. | Treatment of discs large homolog (DLG) related diseases by inhibition of natural antisense transcript to DLG |
CN103210086B (en) | 2010-10-06 | 2017-06-09 | 库尔纳公司 | NEU4 relevant diseases are treated by suppressing the natural antisense transcript of sialidase 4 (NEU4) |
WO2012052391A1 (en) | 2010-10-19 | 2012-04-26 | Glaxo Group Limited | Polypeptide with jmjd3 catalytic activity |
EP2630241B1 (en) | 2010-10-22 | 2018-10-17 | CuRNA, Inc. | Treatment of alpha-l-iduronidase (idua) related diseases by inhibition of natural antisense transcript to idua |
WO2012068340A2 (en) | 2010-11-18 | 2012-05-24 | Opko Curna Llc | Antagonat compositions and methods of use |
WO2012074855A2 (en) | 2010-11-22 | 2012-06-07 | The Regents Of The University Of California | Methods of identifying a cellular nascent rna transcript |
KR102010598B1 (en) | 2010-11-23 | 2019-08-13 | 큐알엔에이, 인크. | Treatment of nanog related diseases by inhibition of natural antisense transcript to nanog |
KR102160721B1 (en) | 2010-12-22 | 2020-09-29 | 페이트 세러퓨틱스, 인코포레이티드 | Cell culture platform for single cell sorting and enhanced reprogramming of iPSCs |
GB201101621D0 (en) | 2011-01-31 | 2011-03-16 | Olink Ab | Method and product |
WO2012109495A1 (en) | 2011-02-09 | 2012-08-16 | Metabolic Solutions Development Company, Llc | Cellular targets of thiazolidinediones |
EP3502236B1 (en) | 2011-02-18 | 2023-08-23 | The Scripps Research Institute | Directed differentiation of oligodendrocyte precursor cells to a myelinating cell fate |
WO2012119989A2 (en) | 2011-03-04 | 2012-09-13 | Oryzon Genomics, S.A. | Methods and antibodies for the diagnosis and treatment of cancer |
US20120301904A1 (en) | 2011-04-26 | 2012-11-29 | Prosetta Antiviral, Inc | Multiprotein assemblies |
US9561274B2 (en) | 2011-06-07 | 2017-02-07 | University Of Hawaii | Treatment and prevention of cancer with HMGB1 antagonists |
US9244074B2 (en) | 2011-06-07 | 2016-01-26 | University Of Hawaii | Biomarker of asbestos exposure and mesothelioma |
CA2838588C (en) | 2011-06-09 | 2021-09-14 | Curna, Inc. | Treatment of frataxin (fxn) related diseases by inhibition of natural antisense transcript to fxn |
EP2738255B1 (en) | 2011-07-29 | 2016-11-23 | Tokushima University | Erap1-derived peptide and use thereof |
AU2012341028C1 (en) | 2011-09-02 | 2017-10-19 | Mount Sinai School Of Medicine | Substituted pyrazolo[3,4-D]pyrimidines and uses thereof |
MX365525B (en) | 2011-09-06 | 2019-06-06 | Curna Inc | TREATMENT OF DISEASES RELATED TO ALPHA SUBUNITS OF SODIUM CHANNELS, VOLTAGE-GATED (SCNxA) WITH SMALL MOLECULES. |
EP2790687B1 (en) | 2011-12-16 | 2018-08-29 | Poseida Therapeutics, Inc. | Trpc4 modulators for use in the treatment or prevention of pain |
GB201201547D0 (en) | 2012-01-30 | 2012-03-14 | Olink Ab | Method and product |
JP6253596B2 (en) | 2012-02-16 | 2017-12-27 | ザ ペン ステイト リサーチ ファンデーション | Acyl coenzyme A: a method for identifying an inhibitor of expression, function or activity of lysocardiolipin acyltransferase 1 (ALCAT1) |
EP2825209B1 (en) | 2012-03-14 | 2018-08-29 | University of Central Florida Research Foundation, Inc. | Neurofibromatoses therapeutic agents and screening for same |
KR20140136488A (en) | 2012-03-15 | 2014-11-28 | 큐알엔에이, 인크. | Treatment of brain derived neurotrophic factor(bdnf) related diseases by inhibition of natural antisense transcript to bdnf |
CN104379563B (en) | 2012-04-10 | 2018-12-21 | 加利福尼亚大学董事会 | Composition and method for treating cancer |
AU2013252909B2 (en) | 2012-04-24 | 2017-09-28 | University Of Miami | Perforin 2 defense against invasive and multidrug resistant pathogens |
WO2013184645A2 (en) | 2012-06-04 | 2013-12-12 | The Scripps Research Institute | Novel phenyl glyoxal probes |
EP2877211A4 (en) | 2012-07-25 | 2016-02-10 | Salk Inst For Biological Studi | Regulating the interaction between tam ligands and lipid membranes with exposed phosphatidylserine |
CA2922849A1 (en) | 2012-08-31 | 2014-03-06 | Ixchel Pharma, Llc | Agents useful for treating obesity, diabetes and related disorders |
US20150274755A1 (en) * | 2012-09-25 | 2015-10-01 | Shane W. Krska | Compound diversification using late stage functionalization |
SG11201502331RA (en) | 2012-09-26 | 2015-04-29 | Univ California | Modulation of ire1 |
AU2013204200B2 (en) | 2012-10-11 | 2016-10-20 | Brandeis University | Treatment of amyotrophic lateral sclerosis |
WO2014066864A2 (en) | 2012-10-26 | 2014-05-01 | Memorial Sloan-Kettering Cancer Center | Androgen receptor variants and methods for making and using |
US20140120116A1 (en) | 2012-10-26 | 2014-05-01 | The Chinese University Of Hong Kong | Treatment of cancer using smad3 inhibitor |
WO2014124339A2 (en) | 2013-02-07 | 2014-08-14 | The Regents Of The University Of California | Use of translational profiling to identify target molecules for therapeutic treatment |
EP2970926B1 (en) | 2013-03-13 | 2018-01-31 | DSM Nutritional Products AG | Engineering microorganisms |
UY35464A (en) | 2013-03-15 | 2014-10-31 | Araxes Pharma Llc | KRAS G12C COVALENT INHIBITORS. |
US9428537B2 (en) | 2013-03-15 | 2016-08-30 | The Board Of Trustees Of The Leland Stanford Junior University | tRNA derived small RNAs (tsRNAs) involved in cell viability |
CN105392882A (en) | 2013-04-19 | 2016-03-09 | 加利福尼亚大学董事会 | Lone star virus |
TWI659021B (en) | 2013-10-10 | 2019-05-11 | 亞瑞克西斯製藥公司 | Inhibitors of kras g12c |
WO2015057461A2 (en) | 2013-10-18 | 2015-04-23 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Antibodies that specifically bind ataxia telangiectasia-mutated and rad3-related kinase phosphorylated at position 1989 and their use |
WO2015089338A2 (en) | 2013-12-11 | 2015-06-18 | Sloan-Kettering Institute For Cancer Research | Glucocorticoid inhibitors for treatment of prostate cancer |
WO2015089327A1 (en) | 2013-12-11 | 2015-06-18 | Biogen Idec Ma Inc. | Biaryl compounds useful for the treatment of human diseases in oncology, neurology and immunology |
US11268069B2 (en) | 2014-03-04 | 2022-03-08 | Fate Therapeutics, Inc. | Reprogramming methods and cell culture platforms |
WO2015136509A2 (en) | 2014-03-14 | 2015-09-17 | Genesis Theranostix Korlatolt Felelossegu Tarsasag | Diagnostic and therapeutic targets for preeclampsia and closely related complications of pregnancy |
JO3556B1 (en) | 2014-09-18 | 2020-07-05 | Araxes Pharma Llc | Combination therapies for treatment of cancer |
WO2016049568A1 (en) | 2014-09-25 | 2016-03-31 | Araxes Pharma Llc | Methods and compositions for inhibition of ras |
WO2016049524A1 (en) | 2014-09-25 | 2016-03-31 | Araxes Pharma Llc | Inhibitors of kras g12c mutant proteins |
US10227333B2 (en) | 2015-02-11 | 2019-03-12 | Curtana Pharmaceuticals, Inc. | Inhibition of OLIG2 activity |
EA039121B1 (en) | 2015-02-27 | 2021-12-07 | Куртана Фармасьютикалс, Инк. | INHIBITION OF Olig2 ACTIVITY |
KR20180005178A (en) | 2015-04-10 | 2018-01-15 | 아락세스 파마 엘엘씨 | Substituted quinazoline compounds and methods for their use |
WO2016168540A1 (en) | 2015-04-15 | 2016-10-20 | Araxes Pharma Llc | Fused-tricyclic inhibitors of kras and methods of use thereof |
US10196701B2 (en) | 2015-06-01 | 2019-02-05 | The Penn State Research Foundation | Hepatitis B virus capsid assembly |
US10851423B2 (en) | 2015-06-22 | 2020-12-01 | Proteovista Llc | SNP arrays |
EP3314260B1 (en) | 2015-06-24 | 2021-04-21 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods for detecting protein-protein interactions |
US10144724B2 (en) | 2015-07-22 | 2018-12-04 | Araxes Pharma Llc | Substituted quinazoline compounds and methods of use thereof |
EP3356347A1 (en) | 2015-09-28 | 2018-08-08 | Araxes Pharma LLC | Inhibitors of kras g12c mutant proteins |
US10647703B2 (en) | 2015-09-28 | 2020-05-12 | Araxes Pharma Llc | Inhibitors of KRAS G12C mutant proteins |
WO2017058915A1 (en) | 2015-09-28 | 2017-04-06 | Araxes Pharma Llc | Inhibitors of kras g12c mutant proteins |
EP3356339A1 (en) | 2015-09-28 | 2018-08-08 | Araxes Pharma LLC | Inhibitors of kras g12c mutant proteins |
EP3356354A1 (en) | 2015-09-28 | 2018-08-08 | Araxes Pharma LLC | Inhibitors of kras g12c mutant proteins |
EP3356359B1 (en) | 2015-09-28 | 2021-10-20 | Araxes Pharma LLC | Inhibitors of kras g12c mutant proteins |
US10858343B2 (en) | 2015-09-28 | 2020-12-08 | Araxes Pharma Llc | Inhibitors of KRAS G12C mutant proteins |
US11441126B2 (en) | 2015-10-16 | 2022-09-13 | Fate Therapeutics, Inc. | Platform for the induction and maintenance of ground state pluripotency |
EP3364977A4 (en) | 2015-10-19 | 2019-09-04 | Araxes Pharma LLC | Method for screening inhibitors of ras |
EP3377481A1 (en) | 2015-11-16 | 2018-09-26 | Araxes Pharma LLC | 2-substituted quinazoline compounds comprising a substituted heterocyclic group and methods of use thereof |
WO2017100546A1 (en) | 2015-12-09 | 2017-06-15 | Araxes Pharma Llc | Methods for preparation of quinazoline derivatives |
US10822312B2 (en) | 2016-03-30 | 2020-11-03 | Araxes Pharma Llc | Substituted quinazoline compounds and methods of use |
US10646488B2 (en) | 2016-07-13 | 2020-05-12 | Araxes Pharma Llc | Conjugates of cereblon binding compounds and G12C mutant KRAS, HRAS or NRAS protein modulating compounds and methods of use thereof |
WO2018036503A1 (en) | 2016-08-25 | 2018-03-01 | The Chinese University Of Hong Kong | Fecal bacterial markers for colorectal cancer |
US10280172B2 (en) | 2016-09-29 | 2019-05-07 | Araxes Pharma Llc | Inhibitors of KRAS G12C mutant proteins |
CN110312711A (en) | 2016-10-07 | 2019-10-08 | 亚瑞克西斯制药公司 | Heterocyclic compound and its application method as RAS inhibitor |
MX2019007030A (en) | 2016-12-15 | 2020-01-15 | Univ California | Compositions and methods for treating cancer. |
WO2018140512A1 (en) | 2017-01-26 | 2018-08-02 | Araxes Pharma Llc | Fused bicyclic benzoheteroaromatic compounds and methods of use thereof |
US11136308B2 (en) | 2017-01-26 | 2021-10-05 | Araxes Pharma Llc | Substituted quinazoline and quinazolinone compounds and methods of use thereof |
US11358959B2 (en) | 2017-01-26 | 2022-06-14 | Araxes Pharma Llc | Benzothiophene and benzothiazole compounds and methods of use thereof |
EP3573971A1 (en) | 2017-01-26 | 2019-12-04 | Araxes Pharma LLC | 1-(3-(6-(3-hydroxynaphthalen-1-yl)benzofuran-2-yl)azetidin-1yl)prop-2-en-1-one derivatives and similar compounds as kras g12c modulators for treating cancer |
JP7327802B2 (en) | 2017-01-26 | 2023-08-16 | アラクセス ファーマ エルエルシー | Fused hetero-heterobicyclic compounds and methods of use thereof |
EP3596117A4 (en) | 2017-03-17 | 2021-01-13 | The Johns Hopkins University | Targeted epigenetic therapy against distal regulatory element of tgfb2 expression |
EP3601326A4 (en) | 2017-03-20 | 2020-12-16 | The Broad Institute, Inc. | Compounds and methods for regulating insulin secretion |
CN110831933A (en) | 2017-05-25 | 2020-02-21 | 亚瑞克西斯制药公司 | Quinazoline derivatives as modulators of mutated KRAS, HRAS or NRAS |
SG10202113146UA (en) | 2017-05-25 | 2021-12-30 | Araxes Pharma Llc | Covalent inhibitors of kras |
JP2020521741A (en) | 2017-05-25 | 2020-07-27 | アラクセス ファーマ エルエルシー | Compounds for the treatment of cancer and methods of their use |
US11725056B2 (en) | 2017-10-03 | 2023-08-15 | Cedars-Sinai Medical Center | Methods for targeting the immune checkpoint PD1 pathway for treating pulmonary fibrosis |
MX2020006150A (en) | 2017-12-15 | 2020-11-11 | Flagship Pioneering Innovations Vi Llc | Compositions comprising circular polyribonucleotides and uses thereof. |
AU2019373090B2 (en) | 2018-10-31 | 2023-05-25 | The University Of Sydney | Compositions and methods for treating viral infections |
US11325978B2 (en) | 2018-11-06 | 2022-05-10 | The United States Of America, As Represented By The Secretary Of The Department Of Health And Human Services | Compositions and methods for treating beta-globinopathies |
AU2020231349A1 (en) | 2019-03-01 | 2021-09-23 | Flagship Pioneering Innovations Vi, Llc | Compositions, methods, and kits for delivery of polyribonucleotides |
WO2020198403A2 (en) | 2019-03-25 | 2020-10-01 | Flagship Pioneering Innovations Vi, Llc | Compositions comprising modified circular polyribonucleotides and uses thereof |
JP2022537154A (en) | 2019-06-14 | 2022-08-24 | フラッグシップ パイオニアリング イノベーションズ シックス,エルエルシー | circular RNA for cell therapy |
AU2020296190A1 (en) | 2019-06-19 | 2022-01-06 | Flagship Pioneering Innovations Vi, Llc | Methods of dosing circular polyribonucleotides |
TW202142239A (en) | 2020-01-29 | 2021-11-16 | 美商旗艦先鋒創新有限責任公司 | Delivery of compositions comprising circular polyribonucleotides |
US11965162B2 (en) | 2020-04-16 | 2024-04-23 | The Johns Hopkins University | MicroRNA and inhibitors thereof and methods of treatment |
WO2023081167A2 (en) | 2021-11-02 | 2023-05-11 | The Regents Of The University Of California | P-selectin mutants and modulation of integrin-mediated signaling |
WO2023146807A1 (en) | 2022-01-25 | 2023-08-03 | The Regents Of The University Of California | Vegf mutants and modulation of integrin-mediated signaling |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE1200276B (en) * | 1959-10-08 | 1965-09-09 | Bayer Ag | Process for the preparation of carboxylic acid esters of non-reducing sugars |
US3859318A (en) * | 1969-05-19 | 1975-01-07 | Lubrizol Corp | Products produced by post-treating oil-soluble esters of mono- or polycarboxylic acids and polyhydric alcohols with epoxides |
US4005195A (en) * | 1976-02-12 | 1977-01-25 | The Procter & Gamble Company | Compositions for treating hypercholesterolemia |
US4241054A (en) * | 1978-12-08 | 1980-12-23 | The Procter & Gamble Company | Detoxifying lipophilic toxins |
DE2855038A1 (en) * | 1978-12-20 | 1980-07-10 | Bayer Ag | METHOD FOR CLEANING LOW MOLECULAR POLYHYDROXYL COMPOUNDS |
US4631211A (en) * | 1985-03-25 | 1986-12-23 | Scripps Clinic & Research Foundation | Means for sequential solid phase organic synthesis and methods using the same |
NZ215865A (en) * | 1985-04-22 | 1988-10-28 | Commw Serum Lab Commission | Method of determining the active site of a receptor-binding analogue |
US5223409A (en) * | 1988-09-02 | 1993-06-29 | Protein Engineering Corp. | Directed evolution of novel binding proteins |
FR2640627A1 (en) * | 1988-12-16 | 1990-06-22 | Commissariat Energie Atomique | New aldosaminyl fluorides, their preparation and their use for the manufacture of (1->4) bonded oligosaccharide amines. |
US5650489A (en) * | 1990-07-02 | 1997-07-22 | The Arizona Board Of Regents | Random bio-oligomer library, a method of synthesis thereof, and a method of use thereof |
US5770358A (en) * | 1991-09-18 | 1998-06-23 | Affymax Technologies N.V. | Tagged synthetic oligomer libraries |
RU2134693C1 (en) * | 1993-02-23 | 1999-08-20 | Зе Трастис оф Принстон Юниверсити | Methods of forming glycoside bonds, chemical composition, glycoside and glycoside library |
WO1995003315A2 (en) * | 1993-07-21 | 1995-02-02 | Oxford Glycosystems Ltd | Saccharides, their synthesis and use |
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